Antibiotics containing borinic acid complexes and methods of use

ABSTRACT

The structure and preparation of antibiotics incorporating borinic acid complexes are disclosed, especially hydroxyquinoline, imidazole and picolinic acid derivatives, along with compositions of these antibiotics and methods of using the antibiotics and compositions as bactericidal and fungicidal agents as well as therapeutic agents for the treatment of diseases caused by bacteria and fungi.

This application is a continuation-in-part of U.S. application Ser. No.10/740,304, filed 18 Dec. 2003, which claimed priority of U.S.Provisional Applications Ser. No. 60/434,375, filed 18 Dec. 2002, Ser.No. 60/436,095, filed 23 Dec. 2002, and Ser. No. 60/437,849, filed 3Jan. 2003, the disclosures of all of which are hereby incorporated byreference in their entirety.

FIELD OF THE INVENTION

The present invention relates to the field of antibiotics andparticularly antibacterial and antifungal compounds and uses thereof.Methods for preparing and using these antibiotics, and pharmaceuticalcompositions thereof, are also provided.

BACKGROUND OF THE INVENTION

One hallmark of the modern era of medicine has been the decline inmorbidity and mortality associated with bacterial and fungal infections.However, misuse of conventional antibiotics and natural selection of theinfectious bacterial population has resulted in the development ofvarying degrees of drug resistance by most bacterial infectious agentsto most antibiotic agents. In severe cases, such as MRSA(Multidrug-Resistant StaphA), one or only a few antibiotics arecurrently effective. In addition, the existence of immunodeficiencysyndromes results in additional incidence of opportunistic infectionsrequiring intensive antibiotic treatment.

Thus, there continues to be a need in the medical arts for novel, moreeffective, antibiotic compounds, especially for treating bacterialinfections, that are resistant to currently available therapies.

Boron containing compounds have received increasing attention astherapeutic agents over the past few years as technology in organicsynthesis has expanded to include this atom. [Boron Therapeutics on thehorizon, Groziak, M. P.; American Journal of Therapeutics (2001) 8,321-328] The most notable boron containing therapeutic is the boronicacid bortezomib which was recently launched for the treatment ofmultiple myeloma. This breakthrough demonstrates the feasibility ofusing boron containing compounds as pharmaceutical agents. Boroncontaining compounds have been shown to have various biologicalactivities including herbicides [Organic boron compounds as herbicides.Barnsley, G. E.; Eaton, J. K.; Airs, R. S.; (1957), DE 101697819571003], boron neutron capture therapy [Molecular Design and Synthesisof B-10 Carriers for Neutron Capture Therapy. Yamamoto, Y.; Pure Appl.Chem., (1991) 63, 423-426], serine protease inhibition [Borinic acidinhibitors as probes of the factors involved in binding at the activesites of subtilisin Carlsberg and α-chymotrypsin. Simpelkamp, J.; Jones,J. B.; Bioorganic & Medicinal Chemistry Letters, (1992), 2(11), 1391-4],[Design, Synthesis and Biological Evaluation of SelectiveBoron-containing Thrombin Inhibitors. Weinand, A.; Ehrhardt, C.;Metternich, R.; Tapparelli, C.; Bioorganic and Medicinal Chemistry,(1999), 7, 1295-1307], acetylcholinesterase inhibition [New, specificand reversible bifunctional alkylborinic acid inhibitor ofacetylcholinesterase. Koehler, K. A.; Hess, G. P.; Biochemistry (1974),13, 5345-50] and as antibacterial agents [Boron-Containing AntibacterialAgents: Effects on Growth and Morphology of Bacteria Under VariousCulture Conditions. Bailey, P. J.; Cousins, G.; Snow, G. A.; and White,A. J.; Antimicrobial Agents and Chemotherapy, (1980), 17, 549-553]. Theboron containing compounds with antibacterial activity can besub-divided into two main classes, the diazaborinines, which have beenknown since the 1960's, and dithienylborinic acid complexes. This latterclass has been expanded to include many different diarylborinic acidcomplexes with potent antibacterial activity [Preparation ofdiarylborinic acid esters as DNA methyl transferase inhibitors.Benkovic, S. J.; Shapiro, L.; Baker, S. J.; Wahnon, D. C.; Wall, M.;Shier, V. K.; Scott, C. P.; Baboval, J.; PCT Int. Appl. (2002), WO2002044184]. Synthetic developments described in Benkovic et al. enabledcreation of a much more diverse class of unsymmetrical di-substitutedborinic acid complexes not possible before.

BRIEF SUMMARY OF THE INVENTION

In one aspect, the present invention relates to borinate derivatives asantibiotic compounds especially borinic acid complexes, and include suchcompounds as derivatives of hydroxyquinolines, picolinic acids andimidazoles.

The antibiotic compounds are also provided as pharmaceuticalcompositions that can be administered to an animal, most preferably ahuman, for treatment of a disease having a bacterial or fungal etiology,or an opportunistic infection with a bacteria or fungus in an animal,most preferably a human, in an immunologically compromised ordebilitated state of health.

In preferred embodiments, the compounds of the invention are thosehaving the structures given by Formulas 1 or 2, with preferredsubstituents as disclosed herein.

The invention also provides methods for preparing the antibioticcompounds and pharmaceutical compositions thereof, and methods of usingsaid antibiotics therapeutically. Kits and packaged embodiments of theantibiotic compounds and pharmaceutical compositions of the inventionare also contemplated.

The invention also relates to methods of treating infections, preferablybacterial and/or fungal infections, using the antibiotic compoundsdisclosed herein.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 shows the results of HPLC (high performance liquidchromatography) for several picolinic acid derivatives of the invention.

FIG. 2 shows the results of HPLC (high performance liquidchromatography) for several picolinic acid derivatives of the invention.

DETAILED DESCRIPTION OF THE INVENTION

This invention provides antibiotics, and specifically antibacterial andanti-fungal compounds, useful in treating and/or preventing bacterialinfections.

The invention comprises a compound having the structure with formula

-   -   wherein B is boron, O is oxygen    -   wherein R* and R** are each independently selected from        substituted or unsubstituted alkyl (C₁-C₄), substituted or        unsubstituted cycloalkyl (C₃-C₇), substituted or unsubstituted        alkenyl, substituted or unsubstituted alkynyl, substituted or        unsubstituted aralkyl, substituted or unsubstituted phenyl, and        substituted or unsubstituted heteroaryl,    -   and wherein z is 0 or 1 and when z is 1, A is CH, CR¹⁰ or N,    -   and wherein D is N, CH, or CR¹²,    -   and wherein E is H, OH, alkoxy or 2-(morpholino)ethoxy, CO₂H or        CO₂alkyl    -   and wherein m=0-2,    -   and wherein r is 1 or 2, and wherein when r is 1, G is ═O        (double-bonded oxygen) and when r is 2, each G is independently        H, methyl, ethyl or propyl,    -   wherein R¹² is selected from (CH₂)_(k)OH (where k=1, 2 or 3),        CH₂NH₂, CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H, CO₂alkyl, CONH₂, OH,        alkoxy, aryloxy, SH, S-alkyl, S-aryl, SO₂N(alkyl)₂, SO₂NHalkyl,        SO₂NH₂, SO₂alkyl, SO₃H, SCF₃, CN, halogen, CF₃, NO₂, NH₂,        2*-amino, 3*-amino, NH₂SO₂ and CONH₂,    -   and wherein J is CR¹⁰ or N    -   and wherein R⁹, R¹⁰ and R¹¹ are each independently selected from        the group consisting of hydrogen, alkyl, cycloalkyl, (CH₂)_(n)OH        (n=2 to 3), CH₂NH₂, CH₂NHalkyl, CH₂N(alkyl)₂, halogen, CHO,        CH═NOH, CO₂H, CO₂-alkyl, S-alkyl, SO₂-alkyl, S-aryl,        SO₂N(alkyl)₂, SO₂NHalkyl, SO₂NH₂, NH₂, alkoxy, CF₃, SCF₃, NO₂,        SO₃H and OH,    -   including salts thereof.

In preferred embodiments of formula 1 or 2, R* and/or R** are the sameor are different, preferably wherein one of R* and R** is a substitutedor unsubstituted alkyl (C₁-C₄) or R* and R** are each a substituted orunsubstituted alkyl (C₁-C₄).

In a preferred embodiment of formulas 1 or 2, R* and/or R** are the sameor are different, preferably wherein one of R* and R** is a substitutedor unsubstituted cycloalkyl (C₃-C₇) or R* and R** are each a substitutedor unsubstituted cycloalkyl (C₃-C₇).

In a preferred embodiment of formulas 1 or 2, R* and/or R** are the sameor are different, preferably wherein one of R* and R** is a substitutedor unsubstituted alkenyl or R* and R** are each a substituted orunsubstituted alkenyl. In a further preferred embodiment thereof, thealkenyl has the structure

-   -   wherein R¹, R², and R³ are each independently selected from the        group consisting of hydrogen, alkyl, aryl, cycloalkyl,        substituted aryl, aralkyl, substituted aralkyl, (CH₂)_(k)OH        (where k=1, 2 or 3), CH₂NH₂, CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H,        CO₂alkyl, CONH₂, S-alkyl, S-aryl, SO₂alkyl, SO₂N(alkyl)₂,        SO₂NHalkyl, SO₂NH₂, SO₃H, SCF₃, CN, halogen, CF₃ and NO₂.

In a preferred embodiment of formulas 1 or 2, R* and/or R** are the sameor are different, preferably wherein one of R* and R** is a substitutedor unsubstituted alkynyl or R* and R** are each a substituted orunsubstituted alkynyl. In a further preferred embodiment thereof, thealkynyl has the structure

-   -   wherein R¹ is selected from the group consisting of hydrogen,        alkyl, cycloalkyl, aryl, substituted aryl, aralkyl, substituted        aralkyl, (CH₂)_(k)OH (where k=1, 2 or 3), CH₂NH₂, CH₂NH-alkyl,        CH₂N(alkyl)₂, CO₂H, CO₂alkyl, CONH₂, S-alkyl, S-aryl, SO₂alkyl,        SO₂N(alkyl)₂, SO₂NHalkyl, SO₂NH₂, SO₃H, SCF₃, CN, halogen, CF₃        and NO₂.

In a preferred embodiment of formulas 1 or 2, R* and/or R** are the sameor are different, preferably wherein one of R* and R** is a substitutedor unsubstituted phenyl or R* and R** are each a substituted orunsubstituted phenyl but excluding compounds of formula 1 wherein z is1, A is CR¹⁰, D is CR¹², J is CR¹⁰ and excluding compounds of formula 2wherein the combination of substituents is such that z is 1, A is CR¹⁰,D is CR¹², m is 2, and G is H or methyl or ethyl. In a separateembodiment of the foregoing, G is also not propyl. However, in specificembodiments such excluded compounds, although not being claimed asnovel, may find use in one or more of the methods of the invention,preferably for treatment against infection, most preferably in treatmentagainst fungal infection. In a preferred embodiment, only novelcompounds of the invention are contemplated for such uses.

The novel compounds of the invention do not include quinaldinederivatives, such as 2-methylquinoline, wherein R⁹ is a methyl, A_(z) isCH, D is CH, J is CH and R¹¹ is hydrogen. However, such compounds may beuseful in the methods of the invention.

A preferred embodiment is a compound of Formula 2 wherein R* and R** areeach other than a phenyl or substituted phenyl.

Another preferred embodiment is a compound of Formula 2 wherein one ofR* or R** is benzyl or substituted benzyl.

An additional preferred embodiment is a compound of Formula 2 wherein ris 1, G is ═O, m is 0 and E is OH.

A preferred embodiment is also a compound of Formula 2 wherein z is 1and R⁹ is selected from alkyl (greater than C₄), (CH₂)_(n)OH (n=1, 2 or3), CH₂NH₂, CH₂NHalkyl, CH₂N(alkyl)₂, CHO, CH═NOH, CO₂H, CO₂-alkyl,S-alkyl, SO₂-alkyl, S-aryl, alkoxy (greater than C₄), SCF₃, and NO₂.

In one preferred embodiment the compound has the structure of Formula 2wherein z is 1 and R¹⁰ is selected from alkyl (greater than C₄),(CH₂)_(n)OH (n=1, 2 or 3), CH₂NH₂, CH₂NHalkyl, CH₂N(alkyl)₂, CHO,CH═NOH, CO₂H, CO₂-alkyl, S-alkyl, SO₂-alkyl, S-aryl, alkoxy (greaterthan C₄), SCF₃, and NO₂.

In another preferred embodiment the compound has the structure ofFormula 2 wherein z is 1 and D is CR¹² wherein R¹² is selected from(CH₂)_(k)OH (where k=1, 2 or 3), CH₂NH₂, CH₂NH-alkyl, CH₂N(alkyl)₂,CO₂H, CO₂alkyl, CONH₂, OH, alkoxy (greater than C₄), aryloxy, SH,S-alkyl, S-aryl, SO₂alkyl, SO₃H, SCF₃, CN, NO₂, NH₂SO₂ and CONH₂.

In an additional preferred embodiment the compound has the structure ofFormula 2 wherein z is 1, E is N-(morpholinyl)ethoxy or alkoxy greaterthan C₄.

Other preferred embodiments are compounds having the structure ofFormula 2 wherein A or D is nitrogen, or wherein m is 2.

In another preferred embodiment, the compound has the structure ofFormula 2 wherein one of R* or R** is substituted phenyl substitutedwith 1 to 5 substituents each of which is independently selected fromalkyl (greater than C₆), aryl, substituted aryl, benzyl, substitutedbenzyl, (CH₂)_(k)OH (where k=1, 2 or 3), CH₂NH₂, CH₂NH-alkyl,CH₂N(alkyl)₂, CO₂H, CO₂alkyl, CONH₂, CONHalkyl, CON(alkyl)₂, OH, alkoxy(greater than C₆), aryloxy, SH, S-alkyl, S-aryl, SO₂alkyl, SO₃H, SCF₃,CN, NO₂, NH₂, 2^(o)-amino, 3^(o)-amino, NH₂SO₂, OCH₂CH₂NH₂,OCH₂CH₂NHalkyl, OCH₂CH₂N(alkyl)₂, oxazolidin-2-yl, and alkyl substitutedoxazolidin-2-yl.

In a further preferred embodiment thereof, the phenyl has the structure

-   -   wherein R⁴, R⁵, R⁶, R⁷ and R⁸ are each independently selected        from the group consisting of hydrogen, alkyl, cycloalkyl aryl,        substituted aryl, aralkyl, substituted aralkyl, (CH₂)_(k)OH        (where k=1, 2 or 3), CH₂NH₂, CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H,        CO₂alkyl, CONH₂, CONHalkyl, CON(alkyl)₂, OH, alkoxy, aryloxy,        SH, S-alkyl, S-aryl, SO₂alkyl, SO₂N(alkyl)₂, SO₂NHalkyl, SO₂NH₂,        SO₃H, SCF₃, CN, halogen, CF₃, NO₂, NH₂, 2^(o)-amino,        3^(o)-amino, NH₂SO₂, OCH₂CH₂NH₂, OCH₂CH₂NHalkyl,        OCH₂CH₂N(alkyl)₂, oxazolidin-2-yl, or alkyl substituted        oxazolidin-2-yl.

One highly preferred embodiment is a compound of formula 1 wherein R* is3-fluorophenyl, R** is 4-chlorophenyl, R⁹ is H, R¹¹ is H, A_(z) is CH, Dis CH, J is CH and may be called (3-fluorophenyl)(4-chlorophenyl)borinicacid 8-hydroxyquinoline ester.

Another preferred embodiment is a compound of formula 1 wherein R* andR** are each 3-(4,4-dimethyloxazolidin-2-yl)phenyl, R⁹ is H, R¹¹ is H,A_(z) is CH, D is CH, J is CH and may be calledbis(3-(4,4-dimethyloxazolidin-2-yl)phenyl)borinic acid8-hydroxyquinoline ester.

An additional preferred embodiment is a compound of formula 1 wherein R*is 3-fluorophenyl, R** is cyclopropyl, R⁹ is H, R¹¹ is H, A_(z) is CH, Dis CH, J is CH and referred to as (3-fluorophenyl)(cyclopropyl)borinicacid 8-hydroxyquinoline ester.

A highly preferred embodiment is a compound of formula 1 wherein R* is4-(N,N-dimethyl)-aminomethylphenyl, R** is 4-cyanophenyl, R⁹ is H, R¹¹is H, A_(z) is CH, D is CH, J is CH and is referred to as(4-(N,N-dimethyl)-aminomethylphenyl)(4-cyanophenyl)borinic acid8-hydroxyquinoline ester.

Another highly preferred embodiment is a compound of formula 2 whereinR* is the same as R** which is 3-chloro4-methylphenyl, R⁹ is H, R¹¹ isH, A_(z) is CH, D is CH and E is OH, m=0, r is 1, G is ═O (double bondedoxygen) and is referred to as bis(3-chloro4-methylphenyl)borinic acid3-hydroxypicolinate ester.

A further highly preferred embodiment is a compound of formula 2 whereinR* is the same as R** which is 2-methyl4-chlorophenyl, R⁹ is H, R¹¹ isH, A_(z) is CH, D is CH and E is OH, m=0, r is 1, G is ═O (double bondedoxygen) and is referred to as bis(2-methyl4-chlorophenyl)borinic acid3-hydroxypicolinate ester.

In a preferred embodiment of formula 1 or 2, R* and/or R** are the sameor are different, preferably wherein one of R* and R** is a substitutedor unsubstituted benzyl or R* and R** are each a substituted orunsubstituted benzyl. In a further preferred embodiment thereof, thebenzyl has the structure

-   -   wherein R⁴, R⁵, R⁶, R⁷ and R⁸ are each independently selected        from the group consisting of alkyl, cycloalkyl, aryl,        substituted aryl, aralkyl, substituted aralkyl, (CH₂)_(k)OH        (where k=1, 2 or 3), CH₂NH₂, CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H,        CO₂alkyl, CONH₂, CONHalkyl, CON(alkyl)₂, OH, alkoxy, aryloxy,        SH, S-alkyl, S-aryl, SO₂alkyl, SO₂N(alkyl)₂, SO₂NHalkyl, SO₂NH₂,        SO₃H, SCF₃, CN, halogen, CF₃, NO₂, NH₂, 2^(o)-amino,        3^(o)-amino, NH₂SO₂, OCH₂CH₂NH₂, OCH₂CH₂NHalkyl,        OCH₂CH₂N(alkyl)₂, oxazolidin-2-yl, or alkyl substituted        oxazolidin-2-yl.

One preferred embodiment is a compound of formula 1 or 2, R* and/or R**are the same or are different, preferably wherein one of R* and R** is asubstituted or unsubstituted heterocycle or R* and R** are each asubstituted or unsubstituted heterocycle. In a further preferredembodiment thereof, the heterocycle has the structure

-   -   wherein X═CH═CH, N═CH, NR¹³ (wherein R¹³═H, alkyl, aryl or        aralkyl), O, or S    -   and wherein Y═CH or N    -   and wherein R¹, R², and R³ are each independently selected from        the group consisting of hydrogen, alkyl, cycloalkyl, aryl,        substituted aryl, aralkyl, substituted aralkyl, (CH₂)_(k)OH        (where k=1, 2 or 3), CH₂NH₂, CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H,        CO₂alkyl, CONH₂, S-alkyl, S-aryl, SO₂alkyl, SO₂N(alkyl)₂,        SO₂NHalkyl, SO₂NH₂, SO₃H, SCF₃, CN, halogen, CF₃, NO₂,        oxazolidin-2-yl, or alkyl substituted oxazolidin-2-yl.

A highly preferred embodiment is a compound of formula 1 wherein R* ispyrid-3-yl, R** is 4-chlorophenyl, R⁹ is H, R¹¹ is H, A_(z) is CH, D isCH, and J is CH (named (pyrid-3-yl)(4-chlorophenyl)borinic acid8-hydroxyquinoline ester).

A highly preferred embodiment is a compound of formula 1 wherein R* is5-cyanopyrid-3-yl, R** is vinyl, R⁹ is H, R¹¹ is H, A_(z) is CH, D isCH, and J is CH (named (5-cyanopyrid-3-yl)(vinyl)borinic acid8-hydroxyquinoline ester).

One preferred embodiment is a compound of formula 1 wherein R⁹ is H, R¹¹is H, A_(z) is CH, D is CH, and J is CH.

Another preferred embodiment is a compound of formula 2 wherein R⁹ is H,R¹¹ is H, A_(z) is CH, D is CH and E is OH, m=0, r is 1, and G is ═O(double bonded oxygen).

The structures of the invention also permit solvent interactions thatmay afford structures (such as Formulas 1 B and 2B) that include atomsderived from the solvent encountered by the compounds of the inventionduring synthetic procedures and therapeutic uses. Thus, such solventstructures can especially insinuate themselves into at least some of thecompounds of the invention, especially between the boron and nitrogenatoms, to increase the ring size of such compounds by one or two atoms.For example, where the boron ring of a structure of the inventioncomprises 5 atoms, including, for example, the boron, a nitrogen, anoxygen and 2 carbons, insinuation of a solvent atom between the boronand nitrogen would afford a 7 membered ring. In one example, use ofhydroxyl and amino solvents may afford structures containing an oxygenor nitrogen between the ring boron and nitrogen atoms to increase thesize of the ring. Such structures are expressly contemplated by thepresent invention, preferably where R*** is H or alkyl

As used herein, the following terms have the stated meaning:

By “alkyl”, “lower alkyl”, and “C₁-C₆ alkyl” in the present invention ismeant straight or branched chain alkyl groups having 1-6 carbon atoms,such as, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl,tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl,3-hexyl, and 3-methylpentyl.

By “alkoxy”, “lower alkoxy”, and “C₁-C₆ alkoxy” in the present inventionis meant straight or branched chain alkoxy groups having 1-6 carbonatoms, such as, for example, methoxy, ethoxy, propoxy, isopropoxy,n-butoxy, sec-butoxy, tert-butoxy, pentoxy, 2-pentyl, isopentoxy,neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy, and 3-methylpentoxy.

By the term “halogen” in the present invention is meant fluorine,bromine, chlorine, and iodine.

By “cycloalkyl”, e.g., C₃-C₇ cycloalkyl, in the present invention ismeant cycloalkyl groups having 3-7 atoms such as, for examplecyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. InC₃-C₇ cycloalkyl groups, preferably in C₅-C₇ cycloalkyl groups, one ortwo of the carbon atoms forming the ring can be replaced with a heteroatom, such as sulfur, oxygen or nitrogen. Examples of such groups arepiperidinyl, piperazinyl, morpholinyl, pyrrolidinyl, imidazolidinyl,oxazolidinyl, perhydroazepinyl, perhydrooxazapinyl, oxepanyl, andperhydrooxepanyl. C₃ and C₄ cycloalkyl groups having a member replacedby nitrogen or oxygen include aziridinyl, azetidinyl, oxetanyl, andoxiranyl.

By “aryl” is meant an aromatic carbocyclic group having a single ring(e.g., phenyl), multiple rings (e.g., biphenyl), or multiple condensedrings in which at least one is aromatic, (e.g.,1,2,3,4-tetrahydronaphthyl, naphthyl, anthryl, or phenanthryl), which isoptionally mono-, di-, or trisubstituted with, e.g., halogen, loweralkyl, lower alkoxy, lower alkylthio, trifluoromethyl, lower acyloxy,aryl, heteroaryl, and hydroxy. Preferred aryl groups include phenyl andnaphthyl, each of which is optionally substituted as defined herein.

By “heteroaryl” is meant one or more aromatic ring systems of 5-, 6-, or7-membered rings containing at least one and up to four heteroatomsselected from nitrogen, oxygen, or sulfur. Such heteroaryl groupsinclude, for example, thienyl, furanyl, thiazolyl, imidazolyl,(is)oxazolyl, pyridyl, pyrimidinyl, (iso)quinolinyl, napthyridinyl,benzimidazolyl, and benzoxazolyl. Preferred heteroaryls are thiazolyl,pyrimidinyl, preferably pyrimidin-2-yl, and pyridyl. Other preferredheteroaryl groups include 1-imidazolyl, 2-thienyl, 1-(or 2-)quinolinyl,1-(or 2-)isoquinolinyl, 1-(or 2-)tetrahydroisoquinolinyl, 2-(or3-)furanyl and 2- tetrahydro-furanyl.

By “ligand” is meant a nitrogen-containing aromatic system which iscapable of forming a dative bond with the Lewis acidic boron center,while appended as a borinate ester moiety. Such ligands are known tothose trained in the arts. Examples are shown in the structures below.

The compounds of the present invention have been implicated in theinhibition of key microbial enzymes, such as bacterial DNAmethyltransferase. Many of the compounds disclosed herein are selectiveinhibitors of methyltransferases in microbes, while not inhibitory formethyltransferases in mammals. However, the anti-bacterial andanti-fungal activity of the compounds of the invention is not limited tothose with said enzyme inhibitory activity, nor is the latter effectnecessarily essential to said therapeutic activity.

The invention also provides embodiments of the compounds disclosedherein as pharmaceutical compositions. The pharmaceutical compositionsof the present invention can be manufactured in a manner that is itselfknown, e.g., by means of a conventional mixing, dissolving, granulating,dragee-making, levigating, emulsifying, encapsulating, entrapping orlyophilizing processes.

Pharmaceutical compositions for use in accordance with the presentinvention thus can be formulated in conventional manner using one ormore physiologically acceptable carriers comprising excipients andauxiliaries that facilitate processing of the active compounds intopreparations that can be used pharmaceutically. Proper formulation isdependent upon the route of administration chosen.

Non-toxic pharmaceutical salts include salts of acids such ashydrochloric, phosphoric, hydrobromic, sulfuric, sulfinic, formic,toluenesulfonic, methanesulfonic, hydroxyethanesulfonic, nitric,benzoic, citric, tartaric, maleic, fumaric hydroiodic, alkanoic such asacetic, HOOC—(CH₂)_(n)—CH₃ where n is 0-4, and the like. Non-toxicpharmaceutical base addition salts include salts of bases such assodium, potassium, calcium, ammonium, and functional equivalents. Thoseskilled in the art will recognize a wide variety of non-toxicpharmaceutically acceptable addition salts.

For injection, the compounds of the invention can be formulated inappropriate aqueous solutions, such as physiologically compatiblebuffers such as Hanks's solution, Ringer's solution, or physiologicalsaline buffer. For transmucosal and transcutaneous administration,penetrants appropriate to the barrier to be permeated are used in theformulation. Such penetrants are generally known in the art.

For oral administration, the compounds can be formulated readily bycombining the active compounds with pharmaceutically acceptable carrierswell known in the art. Such carriers enable the compounds of theinvention to be formulated as tablets, pills, capsules, liquids, gels,syrups, slurries, suspensions and the like, for oral ingestion by apatient to be treated. Pharmaceutical preparations for oral use can beobtained with solid excipient, optionally grinding a resulting mixture,and processing the mixture of granules, after adding suitableauxiliaries, if desired, to obtain tablets. Suitable excipients are, inparticular, fillers such as sugars, including lactose, sucrose,mannitol, or sorbitol; cellulose preparations such as, for example,maize starch, wheat starch, rice starch, potato starch, gelatin, gumtragacanth, methyl cellulose, hydroxypropylmethylcellulose, sodiumcarboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired,disintegrating agents can be added, such as the cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodiumalginate.

Pharmaceutical preparations that can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, the active compounds can be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols. In addition, stabilizers can be added. All formulations fororal administration should be in dosages suitable for suchadministration. For buccal administration, the compositions can take theform of tablets or lozenges formulated in conventional manner.

For administration by inhalation, the compounds for use according to thepresent invention are conveniently delivered in the form of an aerosolspray presentation from pressurized packs or a nebuliser, with the useof a suitable propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In the case of a pressurized aerosol the dosage unitcan be determined by providing a valve to deliver a metered amount.Capsules and cartridges of e.g., gelatin for use in an inhaler, can beformulated containing a powder mix of the compound and a suitable powderbase such as lactose or starch.

The compounds can be formulated for parenteral administration byinjection, e.g., by bolus injection or continuous infusion. Formulationsfor injection can be presented in unit dosage form, e.g., in ampoules orin multi-dose containers, with an added preservative. The compositionscan take such forms as suspensions, solutions or emulsions in oily oraqueous vehicles, and can contain formulatory agents such as suspending,stabilizing and/or dispersing agents.

Pharmaceutical formulations for parenteral administration includeaqueous solutions of the active compounds in water-soluble form.Additionally, suspensions of the active compounds can be prepared asappropriate oily injection suspensions. Suitable lipophilic solvents orvehicles include fatty oils such as sesame oil, or synthetic fatty acidesters, such as ethyl oleate or triglycerides, or liposomes. Aqueousinjection suspensions can contain substances that increase the viscosityof the suspension, such as sodium carboxymethyl cellulose, sorbitol, ordextran. Optionally, the suspension can also contain suitablestabilizers or agents that increase the solubility of the compounds toallow for the preparation of highly concentrated solutions.Alternatively, the active ingredient can be in powder form forconstitution with a suitable vehicle, e.g., sterile pyrogen-free water,before use. The compounds can also be formulated in rectal compositionssuch as suppositories or retention enemas, e.g., containing conventionalsuppository bases such as cocoa butter or other glycerides.

In addition to the formulations described previously, the compounds canalso be formulated as a depot preparation. Such long acting formulationscan be administered by implantation (for example subcutaneously orintramuscularly) or by intramuscular injection. Thus, for example, thecompounds can be formulated with suitable polymeric or hydrophobicmaterials (for example as an emulsion in an acceptable oil) or ionexchange resins, or as sparingly soluble derivatives, for example, as asparingly soluble salt.

A pharmaceutical carrier for the hydrophobic compounds of the inventionis a cosolvent system comprising benzyl alcohol, a nonpolar surfactant,a water-miscible organic polymer, and an aqueous phase. The cosolventsystem can be the VPD co-solvent system. VPD is a solution of 3% w/vbenzyl alcohol, 8% w/v of the nonpolar surfactant polysorbate 80, and65% w/v polyethylene glycol 300, made up to volume in absolute ethanol.The VPD co-solvent system (VPD:5W) consists of VPD diluted 1:1 with a 5%dextrose in water solution. This co-solvent system dissolves hydrophobiccompounds well, and itself produces low toxicity upon systemicadministration. Naturally, the proportions of a co-solvent system can bevaried considerably without destroying its solubility and toxicitycharacteristics. Furthermore, the identity of the co-solvent componentscan be varied: for example, other low-toxicity nonpolar surfactants canbe used instead of polysorbate 80; the fraction size of polyethyleneglycol can be varied; other biocompatible polymers can replacepolyethylene glycol, e.g. polyvinyl pyrrolidone; and other sugars orpolysaccharides can substitute for dextrose.

Alternatively, other delivery systems for hydrophobic pharmaceuticalcompounds can be employed. Liposomes and emulsions are well knownexamples of delivery vehicles or carriers for hydrophobic drugs. Certainorganic solvents such as dimethyl sulfoxide also can be employed,although usually at the cost of greater toxicity. Additionally, thecompounds can be delivered using a sustained-release system, such assemipermeable matrices of solid hydrophobic polymers containing thetherapeutic agent. Various sustained-release materials have beenestablished and are well known by those skilled in the art.Sustained-release capsules can, depending on their chemical nature,release the compounds for a few weeks up to over 100 days. Depending onthe chemical nature and the biological stability of the therapeuticreagent, additional strategies for protein and nucleic acidstabilization can be employed.

The pharmaceutical compositions also can comprise suitable solid or gelphase carriers or excipients. Examples of such carriers or excipientsinclude but are not limited to calcium carbonate, calcium phosphate,various sugars, starches, cellulose derivatives, gelatin, and polymerssuch as polyethylene glycols.

The compounds of the invention can be provided as salts withpharmaceutically compatible counterions. Pharmaceutically compatiblesalts can be formed with many acids, including but not limited tohydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic,phosphoric, hydrobromic, sulfinic, formic, toluenesulfonic,methanesulfonic, nitic, benzoic, citric, tartaric, maleic, hydroiodic,alkanoic such as acetic, HOOC—(CH₂)_(n)—CH₃ where n is 0-4, and thelike. Salts tend to be more soluble in aqueous or other protonicsolvents that are the corresponding free base forms. Non-toxicpharmaceutical base addition salts include salts of bases such assodium, potassium, calcium, ammonium, and the like. Those skilled in theart will recognize a wide variety of non-toxic pharmaceuticallyacceptable addition salts.

For topical administration, the compounds can be formulated readily bycombining the active compounds with pharmaceutically acceptable carrierswell known in the art. Such carriers enable the compounds of theinvention to be formulated as gels, slurries, suspensions and ointmentsfor topical applications. If desired, disintegrating agents can beadded, such as the cross-linked polyvinyl pyrrolidone, agar, or alginicacid or a salt thereof such as sodium alginate.

Pharmaceutical compositions of the compounds of the present inventioncan be formulated and administered through a variety of means, includingsystemic, localized, or topical administration. Techniques forformulation and administration can be found in “Remington'sPharmaceutical Sciences,” Mack Publishing Co., Easton, Pa. The mode ofadministration can be selected to maximize delivery to a desired targetsite in the body. Suitable routes of administration can, for example,include oral, rectal, transmucosal, transcutaneous, or intestinaladministration. Parenteral delivery, including intramuscular,subcutaneous, and intramedullary injections, as well as intrathecal,direct intraventricular, intravenous, intraperitoneal, intranasal, orintraocular injections are also contemplated.

Alternatively, one can administer the compound in a local rather thansystemic manner, for example, via injection of the compound directlyinto a specific tissue, often in a depot or sustained releaseformulation.

Pharmaceutical compositions suitable for use in the present inventioninclude compositions wherein the active ingredients are contained in aneffective amount to achieve its intended purpose. More specifically, atherapeutically effective amount means an amount effective to preventdevelopment of or to alleviate the existing symptoms of the subjectbeing treated. Determination of the effective amounts is well within thecapability of those skilled in the art, especially in light of thedetailed disclosure provided herein.

For any compound used in the method of the invention, thetherapeutically effective dose can be estimated initially from cellculture assays, as disclosed herein. For example, a dose can beformulated in animal models to achieve a circulating concentration rangethat includes the EC₅₀ (effective dose for 50% increase) as determinedin cell culture, i.e., the concentration of the test compound whichachieves a half-maximal inhibition of bacterial cell growth. Suchinformation can be used to more accurately determine useful doses inhumans.

It will be understood, however, that the specific dose level for anyparticular patient will depend upon a variety of factors including theactivity of the specific compound employed, the age, body weight,general health, sex, diet, time of administration, route ofadministration, and rate of excretion, drug combination, the severity ofthe particular disease undergoing therapy and the judgment of theprescribing physician.

For administration to non-human animals, the drug or a pharmaceuticalcomposition containing the drug may also be added to the animal feed ordrinking water. It will be convenient to formulate animal feed anddrinking water products with a predetermined dose of the drug so thatthe animal takes in an appropriate quantity of the drug along with itsdiet. It will also be convenient to add a premix containing the drug tothe feed or drinking water approximately immediately prior toconsumption by the animal.

Preferred compounds of the invention will have certain pharmacologicalproperties. Such properties include, but are not limited to, oralbioavailability, low toxicity, low serum protein binding and desirablein vitro and in vivo half-lives. Assays may be used to predict thesedesirable pharmacological properties. Assays used to predictbioavailability include transport across human intestinal cellmonolayers, including Caco-2 cell monolayers. Serum protein binding maybe predicted from albumin binding assays. Such assays are described in areview by Oravcová et al. (1996, J. Chromat B 677: 1-27). Compoundhalf-life is inversely proportional to the frequency of dosage of acompound. In vitro half-lives of compounds may be predicted from assaysof microsomal half-life as described by Kuhnz and Gieschen (DrugMetabolism and Disposition, (1998) volume 26, pages 1120-1127).

Toxicity and therapeutic efficacy of such compounds can be determined bystandard pharmaceutical procedures in cell cultures or experimentalanimals, e.g., for determining the LD₅₀ (the dose lethal to 50% of thepopulation) and the ED₅₀ (the dose therapeutically effective in 50% ofthe population). The dose ratio between toxic and therapeutic effects isthe therapeutic index and it can be expressed as the ratio between LD₅₀and ED₅₀. Compounds that exhibit high therapeutic indices are preferred.The data obtained from these cell culture assays and animal studies canbe used in formulating a range of dosage for use in humans. The dosageof such compounds lies preferably within a range of circulatingconcentrations that include the ED₅₀ with little or no toxicity. Thedosage can vary within this range depending upon the dosage formemployed and the route of administration utilized. The exactformulation, route of administration and dosage can be chosen by theindividual physician in view of the patient's condition. (See, e.g.Fingl et al, 1975, in “The Pharmacological Basis of Therapeutics”, Ch.1,p.1).

Dosage amount and interval can be adjusted individually to provideplasma levels of the active moiety that are sufficient to maintainbacterial cell growth inhibitory effects. Usual patient dosages forsystemic administration range from 100-2000 mg/day. Stated in terms ofpatient body surface areas, usual dosages range from 50-910 mg/m²/day.Usual average plasma levels should be maintained within 0.1-1000 μM. Incases of local administration or selective uptake, the effective localconcentration of the compound cannot be related to plasma concentration.

The compounds of the invention are useful as antibiotics for thetreatment of diseases of both animals and humans, including but notlimited to actinomycosis, anthrax, bacterial dysentery, botulism,brucellosis, cellulitis, cholera, conjunctivitis, cystitis, diphtheria,bacterial endocarditis, epiglottitis, gastroenteritis, glanders,gonorrhea, Legionnaire's disease, leptospirosis, bacterial meningitis,plague, bacterial pneumonia, puerperal sepsis, rheumatic fever, RockyMountain spotted fever, scarlet fever, streptococcal pharyngitis,syphilis, tetanus, tuberculosis, tularemia, typhoid fever, typhus, andpertussis.

The disclosures in this application of all articles and references,including patents and patent applications, are incorporated herein byreference in their entirety.

The compounds of the invention comprise a novel class of broad-spectrumantibiotics. Medically-important bacterial species that provideappropriate targets for the antibacterial activity of the inhibitors ofthe invention include gram-positive bacteria, including cocci such asStaphylococcus species and Streptococcus species; acid-fast bacterium,including Mycobacterium species; bacilli, including Bacillus species,Corynebacterium species and Clostridium species; filamentous bacteria,including Actinomyces species and Streptomyces species; gram-negativebacteria, including cocci such as Neisseria species and Acinetobacterspecies; bacilli, such as Pseudomonas species, Brucella species,Agrobacterium species, Bordetella species, Escherichia species, Shigellaspecies, Yersinia species, Salmonella species, Klebsiella species,Enterobacter species, Haemophilus species, Pasteurella species, andStreptobacillus species; spirochetal species, Campylobacter species,Vibrio species; and intracellular bacteria including Rickettsiae speciesand Chlamydia species.

Specific bacterial species that are targets for the antibiotics of theinvention include Staphylococcus aureus; Staphylococcus epidermidis,Staphylococcus saprophyticus; Streptococcus pyogenes; Streptococcusagalactiae; Streptococcus pneumoniae; Enterococcus faecalis;Enterococcus faecium; Bacillus anthracis; Mycobacterium avium,Mycobacterium tuberculosis, Acinetobacter baumannii; Corynebacteriumdiphtheria; Clostridium perfringens; Clostridium botulinum; Clostridiumtetani; Neisseria gonorrhoeae; Neisseria meningitidis; Pseudomonasaeruginosa; Legionella pneumophila; Escherichia coli; Yersinia pestis;Haemophilus influenzae; Helicobacter pylori; Campylobacter fetus;Campylobacter jejuni; Vibrio cholerae; Vibrio parahemolyticus; Trepomenapallidum; Actinomyces israelii; Rickettsia prowazekii; Rickettsiarickettsii; Chlamydia trachomatis; Chlamydia psittaci; Brucella abortus;Agrobacterium tumefaciens; and Francisella tularensis.

Medically-important fungal and yeast species that provide appropriatetargets for the antifungal activity of the inhibitors of the inventioninclude Candida albicans, Candida glabrata, Candida krusei, Candidaparapsilosis, Trichophyton mentagrophytes, Microporium canis,Aspergillus species, Cryptococcus neoformans, Blastomyces dermatitidis,Coccidiodes immitis, Histoplasma capsulatum and Paracoccidioidesbrasiliensis.

In carrying out the procedures of the present invention it is of courseto be understood that reference to particular buffers, media, reagents,cells, culture conditions and the like are not intended to be limiting,but are to be read so as to include all related materials that one ofordinary skill in the art would recognize as being of interest or valuein the particular context in which that discussion is presented. Forexample, it is often possible to substitute one buffer system or culturemedium for another and still achieve similar, if not identical, results.Those skilled in the art will have sufficient knowledge of such systemsand methodologies so as to be able, without undue experimentation, tomake such substitutions as will optimally serve their purposes in usingthe methods and procedures disclosed herein.

The invention is described in more detail in the following non-limitingexamples. It is to be understood that these methods and examples in noway limit the invention to the embodiments described herein and thatother embodiments and uses will no doubt suggest themselves to thoseskilled in the art.

The compounds of this invention are evaluated for their antibacterialactivity as per the guidelines and procedures prescribed by the NationalCommittee for Clinical Laboratory Standards (NCCLS) (cf., NCCLS DocumentM7-A3, 1993 —Antimicrobial Susceptibility Testing).

Protocol for MIC Determination

A useful protocol for MIC determination is as follows:

-   -   1. Approximately 2.5 mg of the compounds to be tested was        weighed into cryovials.    -   2. 5 mg/ml stock solutions were made by adding DMSO to the        samples accordingly.    -   3. 256 μg/ml working solutions were made by using the 5 mg/ml        stock solutions and adding sterile distilled water accordingly.    -   4. A Beckman 2000 Automated Workstation was programmed to load        96 well plates with broth and compounds as follows:        -   100 μl of the appropriate broth was added to columns 1-11        -   200 μl of the appropriate broth was added to column 12        -   100 μl of compounds at the 256 μg/ml working solution were            added to column 1 (one compound per row)        -   Two-fold serial dilutions were done from column 1 to 10        -   Column 11 served as the growth control    -   5. The 10 organism panel was plated from stock vials stored at        −80° C. and incubated for 24 hours at 34° C. The organisms were        then sub-cultured and incubated for 24 hours at 34° C.        -   The inoculums were first prepared in sterile distilled water            with a target of 0.09-0.11 absorbance at 620 nm wavelength        -   A {fraction (1/100)} dilution was made into the appropriate            broth        -   100 μl of broth with organism was added to columns 1-11        -   Column 12 served as the blank control    -   6. The completed 96 well plates were incubated for 24 hours at        34° C. The 96 well plates were then read using a Beckman        Automated Plate Reader at 650 nm wavelength. The MIC was        determined through calculations involving the growth control        (column 11) and blank control (column 12).

Protocol for Antifungal In Vitro MIC Determination

A useful protocol for antifungal activity determination is describedbelow.

Preparation

Media is prepared 1-2 weeks before the start of the experiment. Media isstored in the cold room (4° C.) prior to use.

-   -   Sabouraud Dextrose Agar Plates:        -   1. Add 65 g of powdered of Sabouraud Dextrose Agar media            into 1 L of dH₂O with gentle stirring        -   2. Autoclave at 121° C. and 22 psi for 15 minutes        -   3. Allow the media to cool to ˜50° C.        -   4. Pour media into 100×15 mm sterile petri dishes with 20 ml            aliquots    -   RPMI 1640+MOPS Broth:        -   1. Add 1 packet of powdered RPMI media to 1 L of dH₂O (15°            C.-30° C.) with gentle stirring        -   2. Add 2 g of NaHCO₃        -   3. Add 34.5 g of MOPS        -   4. Adjust the pH to 7.0 using NaOH or HCl        -   5. Sterilize with membrane filtration (0.22 micron cellulose            acetate filter)    -   Sterile Saline (0.9%)        -   1. Dissolve 9 g of NaCl to 1 L of dH₂O        -   2. Autoclave at 121° C. and 22 psi for 15 minutes    -   Sterile dH₂O        -   1. Autoclave dH₂O at 121° C. and 22 psi for 15 minutes            Procedure

1. The 10 organism panel is plated from stock vials stored at −80° C.(suspended in broth with 20% glycerol) and incubated at 37° C. for 24hours. The organisms are then sub-cultured and incubated at 37° C. for24 hours. These will be used to prepare fresh inoculums for Step 6.

2. Approximately 2.5 mg of the compounds to be tested are weighed into 2ml cryovials. Fluconazole, Amphotericin B and Itraconazole are tested asreference compounds.

3. 5 mg/ml stock solutions are made by adding DMSO to the samplesaccordingly. Compounds insoluble with vortexing only are sonicated.

4. 256 μg/ml working solutions are made by using the 5 mg/ml stocksolutions and adding sterile distilled water accordingly.

5. 96-well plates are used for MIC determination. Each of the 8 rows canbe used to test a different compound. Compounds are loaded into thefirst column and two-fold dilutions of are made from column 1 to 10.Column 11 is a growth control (no compound) and column 12 is a blankcontrol (no compound or organism). Manual addition of broth andcompounds is performed as follows:

-   -   100 μl of RPMI+MOPS broth is added to columns 1-11    -   200 μl of RPMI+MOPS broth is added to column 12    -   100 μl of compounds at the 256 μg/ml working solution are added        to column 1 (one compound per row)    -   Two-fold serial dilutions are done from column 1 to 10    -   Column 11 serves as the growth control (media+organism only)

6. The sub-cultured organisms are used to prepare fresh inoculums fortesting on the 96-well plates. Each 96-well plate will test a differentorganism.

-   -   Colonies from the sub-cultured organisms (Step 1) are used to        prepare inoculums with sterile saline. The target is adjusted to        70-75% transmittance at 530 nm wavelength using a Novospec II        spectrophotometer.    -   {fraction (1/1000)} dilution is made into RPMI+MOPS broth    -   100 μl of this broth with organism is added to columns 1-11        (column 12 serves as the blank control)    -   7. The completed 96-well plates are incubated at 37° C. for 24        hours. The 96 well plates are then read for absorbance at 650 nm        wavelength using a Biomek Automated Plate Reader.        Calculations

The absorbance readings from the Biomek Automated Plate Reader are usedto determine the percent inhibition for each test well. The formula usedis as follows:% Inhibition=[1−(ABS _(test) −ABS _(blank))/(ABS _(mean growth) −ABS_(blank))]×100%

-   -   ABS_(test): Absorbance of the test well    -   ABS_(blank): Absorbance of the blank well in the same row as the        test well (column 12)    -   ABS_(mean growth): Mean absorbance of the growth control wells        (column 11)        The minimum inhibitory concentration (MIC) is found at the        lowest concentration of compound where percent inhibition is        greater than or equal to 80%.

These procedures were used to obtain the results in the followingtables. Representative microbiological data for the compounds 10 to 123are shown in Tables 1 to 4 as MIC (Minimum Inhibitory Concentration)with the values expressed as micrograms per ml.

Thus, the invention provides antibiotics that are generically calledborinic acid complexes, most preferably derived from disubstitutedborinic acids.

Borinate Complexes

The synthesis of the compounds of the invention is accomplished inseveral formats. Reaction scheme #1 demonstrates the synthesis of theintermediate borinic acids, and their subsequent conversion to thedesired borinic acid complexes. When R* and R** are identical, thereaction of two equivalents of an arylmagnesium halide (or aryllithium)with trialkyl borate, followed by acidic hydrolysis affords the desiredborinic acid 5. When R* and R** are not identical, the reaction of oneequivalent of an arylmagnesium halide (or aryllithium) with appropriatearyl(dialkoxy)borane (4), heteroaryl(dialkoxy)borane oralkyl(dialkoxy)borane (alkoxy group comprised of methoxy, ethoxy,isopropoxy, or propoxy moiety), followed by acidic hydrolysis affordsthe unsymmetrical borinic acids 6 in excellent yields. Where applicable,the reaction of the alkylene esters (3, T=nothing, CH₂, CMe₂) with theappropriate organocerium, organolithium, organomagnesium or equivalentreactant is convenient.

As shown in Scheme 1, the borinic acid complexes are obtained from theprecursor borinic acids by reaction with one equivalent of the desiredheterocyclic ligand in suitable solvents (i.e., ethanol, isopropanol,dioxane, ether, toluene, dimethylformamide, N-methylpyrrolidone, ortetrahydrofuran).

In certain situations, compounds of the invention may contain one ormore asymmetric carbon atoms, so that the compounds can exist indifferent stereoisomeric forms. These compounds can be, for example,racemates or optically active forms. In these situations, the singleenantiomers, i.e., optically active forms, can be obtained by asymmetricsynthesis or by resolution of the racemates. Resolution of the racematescan be accomplished, for example, by conventional methods such ascrystallization in the presence of a resolving agent, or chromatography,using, for example a chiral HPLC column.

Unexpected Stability of Picolinic Acid Complexes

During the preparation of compounds of Formula 2 we found that complexescontaining hydroxymethyl-type borinic esters, e.g.,2-hydroxymethyl-pyridine (R⁹ is H, A is CH, D is CH, E is H, m is 0, ris 2, G is H) were not stable under typical HPLC conditions. Forexample, the 2-hydroxymethylpyridine complexes shown in FIGS. 1A and 1Bboth eluted as two peaks, one with the solvent front and one much later,indicating some manner of degradation had occurred during the HPLCprocess. This posed a serious problem. Instability issues prevent thisclass of compound from being developed as drug candidates. However, itwas subsequently found that picolinic acid complexes (e.g., R⁹ is H, Ais CH, D is CH, E is H, m is 0, r is 1, G is ═O (double bonded oxygen))were stable under identical the same HPLC conditions. For example, thepicolinic acid complexes shown in FIGS. 2A and 2B show the complex toelute as a single peak indicating no breakdown of the complex. Thebiological activities of the picolinic acid class are comparable to orbetter than the hydroxymethyl pyridine complexes making the picolinicacid complexes an ideal drug development candidate.

HPLC Conditions Sample preparation: ˜1 mg/mL dissolved in 100%acetonitrile Column: BetaBasic C18 5 um 150 × 4.6 mm Flow rate: 1 mL/minInjection vol: 10 uL Wavelength: PDA 200-450 nm; 254 nm reported Mobilephase: Solvent A: 0.1% H₃PO₄ in water Solvent B: Acetonitrile GradientMethod: Time (min) % A % B  0 95 5  1 95 5 11 0 100 21 0 100

Representative compounds of the present invention include, but are notlimited to, the compounds disclosed herein and their pharmaceuticallyacceptable acid and base addition salts. In addition, if the compound ofthe invention is obtained as an acid addition salt, the free base can beobtained by basifying a solution of the acid salt. Conversely, if theproduct is a free base, an addition salt, particularly apharmaceutically acceptable addition salt, may be produced by dissolvingthe free base in a suitable organic solvent and treating the solutionwith an acid, in accordance with conventional procedures for preparingacid addition salts from base compounds. In a preferred embodiment, thecompounds of the invention comprise any of compounds 10-123 (Tables 1,2, 3 and 4), and variants thereof. TABLE 1 Antibacterial Profile AgainstSelect Gram-positive and Gram-negative Pathogens S. S. M. S. aureusepidermidis pneumoniae E. faecalis E. catarrhalis ATCC ATCC ATCC ATCCfaecium ATCC Cmp R* R** Ligand 29213 12228 6301 29212 CT-26 25238 103-ClC₆H₄ 3-ClC₆H₄ 8-hydroxyquinoline 1 2 2 32 4 NA 11 4-Me-3-ClC₆H₃4-Me-3-ClC₆H₃ 4-hydroxybenzimidazole 0.125 4 NA 8 8 NA 12 3-ClC₆H₄3-ClC₆H₄ 5-fluoro-8- 0.125 2 2 8 4 2 hydroxyquinoline 13 3-ClC₆H₄3-ClC₆H₄ 5-chloro-8- 0.125 1 1 64 2 0.25 hydroxyquinoline 14 3-ClC₆H₄3-ClC₆H₄ 4-methyl-8- 0.125 1 1 64 4 0.5 hydroxyquinoline 15 2-F-4-ClC₆H₃3-FC₆H₄ 8-hydroxyquinoline 0.125 1 2 16 4 0.5 16 4-Me-3-ClC₆H₃4-Me-3-ClC₆H₃ 2-HO₂C-4-hydroxy-5,7- 0.25 0.5 NA 0.5 0.25 NAdichloroquinoline 17 3-ClC₆H₄ 3-ClC₆H₄ 2-amino-8- 0.25 2 2 8 8 2hydroxyquinoline 18 3-ClC₆H₄ 3-Cl-4-FC₆H₃ 8-hydroxyquinoline 0.25 1 2 84 1 19 3-ClC₆H₄ 3-ClC₆H₄ 5-cyano-8- 0.25 2 4 16 4 0.5 hydroxyquinoline20 3-ClC₆H₄ 3-Cl-5-FC₆H₃ 8-hydroxyquinoline 0.25 1 2 8 4 2 21 3-ClC₆H₄3-FC₆H₄ 5-cyano-8- 0.5 4 2 16 8 0.25 hydroxyquinoline 22 3-ClC₆H₄3-FC₆H₄ 5-nitro-8- 0.5 4 2 64 16 0.12 hydroxyquinoline 23 3-ClC₆H₄3-FC₆H₄ 5-chloro-7-chloro-8- 0.5 16 8 64 16 0.12 hydroxyquinoline 243-ClC₆H₄ 3-FC₆H₄ 5-bromo-7-bromo-8- 0.5 8 8 64 32 0.12 hydroxyquinoline25 3-ClC₆H₄ 3-ClC₆H₄ 2-carboxy-4-hydroxy-8- 0.5 8 2 16 16 2methoxyquinoline 26 2-thienyl Me 8-hydroxyquinoline 0.5 1 NA 4 4 NA 273-NCC₆H₄ 4-Me-3-ClC₆H₃ 8-hydroxyquinoline 0.5 1 1 8 2 1 28 3,4-Cl₂C₆H₃3-FC₆H₄ 8-hydroxyquinoline 0.5 1 2 4 2 1 29 2,4-Cl₂C₆H₃ 3-FC₆H₄8-hydroxyquinoline 0.5 1 2 8 2 0.5 30 3,4-Cl₂C₆H₃ 3,4-Cl₂C₆H₃8-hydroxyquinoline 1 0.5 NA 2 2 NA 31 4-Me-3-ClC₆H₃ 4-Me-3-ClC₆H₃2-carboxy-4- 1 1 NA 2 1 NA hydroxyquinoline 32 3-ClC₆H₄ 3-FC₆H₄8-hydroxyquinoline 1 1 1 16 2 2 33 3-Cl-5-FC₆H₃ 3-MeC₆H₄8-hydroxyquinoline 1 1 1 8 2 2 34 3-ClC₆H₄ 3-FC₆H₄ 5-fluoro-8- 1 2 2 8 41 hydroxyquinoline 35 3-ClC₆H₄ 3-MeSC₆H₄ 5-fluoro-8- 1 2 2 8 4 2hydroxyquinoline 36 3-ClC₆H₄ 2-thienyl 8-hydroxyquinoline 1 1 2 8 2 4 373-Me-4-ClC₆H₃ 3-NCC₆H₄ 8-hydroxyquinoline 1 1 1 8 2 1 38 2-FC₆H₄3-NCC₆H₄ 8-hydroxyquinoline 1 1 2 16 2 1 39 3-ClC₆H₄ 3-NCC₆H₄8-hydroxyquinoline 1 1 1 8 2 2 40 3-NCC₆H₄ Vinyl 8-hydroxyquinoline 1 11 8 2 2 41 3-NCC₆H₄ Ethynyl 8-hydroxyquinoline 1 1 1 4 2 1 42 3-FC₆H₄Ethynyl 8-hydroxyquinoline 1 1 1 8 2 1 43 2-ClC₆H₄ Ethynyl8-hydroxyquinoline 1 1 1 8 2 1 44 Ethynyl Ethynyl 8-hydroxyquinoline 1 11 16 2 0.25 45 3,5-F₂C₆H₃ Ethynyl 8-hydroxyquinoline 1 1 1 8 1 1 463,5-Cl₂C₆H₃ Ethynyl 8-hydroxyquinoline 1 1 1 4 1 1 47 3,4-Cl₂C₆H₃Ethynyl 8-hydroxyquinoline 1 1 1 8 2 1 48 3-Cl-4-FC₆H₃ Ethynyl8-hydroxyquinoline 1 1 1 8 2 2 49 4-ClC₆H₄ 4-ClC₆H₄ 5-chloro-8- 2 1 1 162 0.25 hydroxyquinoline 50 4-ClC₆H₄ 4-ClC₆H₄ 8-hydroxyquinoline 2 2 2 44 NA 51 3-FC₆H₄ 3-FC₆H₄ 8-hydroxyquinoline 2 1 NA 8 2 NA 524-Me-3-ClC₆H₃ 4-Me-3-ClC₆H₃ 8-hydroxyquinoline 2 4 NA 8 16 NA 533-NCC₆H₄ 3-NCC₆H₄ 8-hydroxyquinoline 2 2 NA 64 4 2 54 4-ClC₆H₄4-Cl-3-FC₆H₃ 8-hydroxyquinoline 2 1 2 8 4 1 55 4-Cl-3-FC₆H₃ 4-Cl-3-FC₆H₃8-hydroxyquinoline 2 1 2 4 2 4 56 3-MeC₆H₄ 3,5-Cl₂C₆H₃8-hydroxyquinoline 2 2 2 8 4 4 57 4-ClC₆H₄ 4-FC₆H₄ 5-fluoro-8- 2 2 4 164 1 hydroxyquinoline 58 3-ClC₆H₄ 4-FC₆H₄ 5-fluoro-8- 2 2 2 8 4 0.5hydroxyquinoline 59 3-ClC₆H₄ 4-MeSC₆H₄ 8-hydroxyquinoline 2 1 1 8 4 2 604-ClC₆H₄ 3-MeSC₆H₄ 8-hydroxyquinoline 2 1 2 8 4 2 61 3-ClC₆H₄cyclopropyl 8-hydroxyquinoline 2 1 1 16 2 2 62 4-ClC₆H₄ 3-MeSC₆H₄5-fluoro-8- 2 2 2 8 4 2 hydroxyquinoline 63 4-ClC₆H₄ 4-MeSC₆H₄8-hydroxyquinoline 2 1 2 8 4 1 64 4-ClC₆H₄ 4-MeSC₆H₄ 5-fluoro-8- 2 2 4 88 1 hydroxyquinoline 65 4-ClC₆H₄ 4-Cl-3-HOC₆H₃ 8-hydroxyquinoline 2 2 216 4 4 66 4-ClC₆H₄ 3-FC₆H₄ 4-methyl-8- 2 1 1 64 4 0.5 hydroxyquinoline67 3-ClC₆H₄ 3-(DMISO)C₆H₄ 4-methyl-8- 2 1 1 64 4 0.5 hydroxyquinoline 683-FC₆H₄ 3-(DMISO)C₆H₄ 8-hydroxyquinoline 2 2 16 32 4 0.12 693-(DMISO)C₆H₄ cyclopropyl 8-hydroxyquinoline 2 1 2 64 4 1 70 3-FC₆H₄cyclopropyl 8-hydroxyquinoline 2 1 1 64 2 0.5 71 3-FC₆H₄ 4-NCC₆H₄5-chloro-7-chloro-8- 2 2 8 64 4 0.12 hydroxyquinoline 72 3-(DMISO)C₆H₄3-(DMISO)C₆H₄ 8-hydroxyquinoline 4 2 4 64 4 NA 73 3-(DMISO)C₆H₄ Vinyl8-hydroxyquinoline 2 1 2 64 8 0.25 74 4-FC₆H₄ 4-NCC₆H₄8-hydroxyquinoline 2 1 2 32 2 1 75 3-ClC₆H₄ 3-MeSC₆H₄ 8-hydroxyquinoline2 1 2 64 4 NA 76 4-Me-3-ClC₆H₃ 2-thienyl 8-hydroxyquinoline 2 1 NA 8 4NA 77 3-ClC₆H₄ 2-MeC₆H₄ 8-hydroxyquinoline 2 1 1 8 4 2 78 3-ClC₆H₄2-MeOC₆H₄ 8-hydroxyquinoline 2 1 1 8 2 2 79 3-ClC₆H₄ 2-Me-4-ClC₆H₃8-hydroxyquinoline 2 2 2 8 4 2 80 4-Cl-3-MeC₆H₃ 4-Cl-3-MeC₆H₃8-hydroxyquinoline 2 1 2 4 4 2 81 3-ClC₆H₄ 3-Cl-6-MeOC₆H₃8-hydroxyquinoline 2 1 2 8 4 2 82 3,5-Cl₂C₆H₃ 4- 8-hydroxyquinoline 2 22 8 2 4 (Me₂NC₂H₄)OC₆H₄ 83 4-BrC₆H₄ 4- 8-hydroxyquinoline 2 1 2 4 4 2(Me₂NC₂H₄)OC₆H₄ 84 3-ClC₆H₄ 4-F-3-MeC₆H₃ 8-hydroxyquinoline 2 1 2 8 4 485 3-Me-4-ClC₆H₃ 3-F-4-ClC₆H₃ 8-hydroxyquinoline 2 1 2 4 4 2 86 3-FC₆H₄4-Cl-3-MeC₆H₃ 8-hydroxyquinoline 2 1 2 8 4 2 87 3-FC₆H₄ 3-F-4-ClC₆H₃8-hydroxyquinoline 2 1 2 8 4 1 88 3-Cl-6-FC₆H₃ 3-NCC₆H₄8-hydroxyquinoline 2 2 2 8 2 2 89 2,5-F₂C₆H₃ 3-NCC₆H₄ 8-hydroxyquinoline2 1 1 8 2 2 90 4-F-3-ClC₆H₃ 3-NCC₆H₄ 8-hydroxyquinoline 2 2 1 8 2 2 913-Me-4-ClC₆H₃ 4-NCC₆H₄ 8-hydroxyquinoline 2 1 2 8 2 1 92 2,5-F₂C₆H₃4-NCC₆H₄ 8-hydroxyquinoline 2 1 2 8 4 1 93 3-Cl-6-FC₆H₃ 4-NCC₆H₄8-hydroxyquinoline 2 1 1 8 4 1 94 3-Cl-6-MeOC₆H₃ 4-NCC₆H₄8-hydroxyquinoline 2 1 1 8 4 2 95 4-NCC₆H₄ Ethynyl 8-hydroxyquinoline 21 2 8 2 1 96 4-ClC₆H₄ 3,4-F₂C₆H₃ 8-hydroxyquinoline 2 1 2 8 2 1 974-ClC₆H₄ 4-Me-3-FC₆H₃ 8-hydroxyquinoline 2 1 1 8 2 1 98 4-ClC₆H₄3,5-F₂C₆H₃ 8-hydroxyquinoline 2 1 1 8 4 1 99 3-CF₃-4-ClC₆H₃ 3-FC₆H₄8-hydroxyquinoline 2 1 2 8 2 1 100 4-ClC₆H₄ 3-F-5-CF₃C₆H₃8-hydroxyquinoline 2 1 2 4 2 1 Ciprofloxacin 0.125 0.125 0.5 0.5 640.125 Cloxacillin 0.125 0.25 0.125 16 64 1 Imipenem 0.125 0.125 0.125 164 0.125 Ceftriaxone 2 1 0.125 64 64 0.125 Meropenem 0.06 0.06 2Erythromycin 0.5 0.5 2 Pen G 0.5 16 0.125 1 32 0.125 DMISO =4,4-dimethyloxazolin-2-yl

TABLE 2 Antibacterial Profile Against Select Gram-positive andGram-negative Pathogens S. S. H. S. aureus epidermidis pneumonia E.faecalis E. influenzae ATCC ATCC ATCC ATCC faecium ATCC Cmp R* R**Ligand 29213 12228 6301 29212 CT-26 49766 101 3-ClC₆H₄ 3-ClC₆H₄1-(2-morpholino-4-yl-ethyl)- 0.12 4 16 64 64 4 imidazoleacetate 1023-ClC₆H₄ 3-ClC₆H₄ 2-hydroxyisopropyl-3- 0.5 1 0.25 64 2 64hydroxypyridine 103 4-ClC₆H₄ 4-ClC₆H₄ 2-hydroxyisopropyl-3- 0.25 0.5 0.54 1 64 hydroxypyridine 104 4-Me-3-ClC₆H₃ 4-Me-3-ClC₆H₃2-hydroxymethyl-1N- 0.5 4 NA 16 32 8 benzylimidazole 105 3-ClC₆H₄3-ClC₆H₄ 2-hydroxymethylpyridine 0.125 4 4 32 32 4 106 4-Me-3-ClC₆H₃4-Me-3-ClC₆H₃ 2-pyridylacetic acid 0.5 4 NA 64 64 64 107 4-Me-3-ClC₆H₃4-Me-3-ClC₆H₃ 3-(2-hydroxyethoxy) 0.125 4 NA 16 8 32 picolinic acid 1084-Me-3-ClC₆H₃ 4-Me-3-ClC₆H₃ 3-(N-morpholinylethoxy) 0.25 4 NA 4 2 64picolinic acid 109 4-Me-3-ClC₆H₃ 4-Me-3-ClC₆H₃ 3-(OCH₂CH₂CH₂CO₂H) 1 4 432 16 16 picolinic acid 110 4-Me-3-ClC₆H₃ 4-Me-3-ClC₆H₃3-carboxypicolinic acid 0.125 4 NA 8 8 8 111 4-Me-3-ClC₆H₃ 4-Me-3-ClC₆H₃3-hydroxypicolinic acid 2 1 NA 2 2 64 112 4-Me-3-ClC₆H₃ 4-CH₃C₆H₄3-hydroxypicolinic acid 4 2 NA 4 8 64 113 4-Me-3-ClC₆H₃ Phenylethyl3-hydroxypicolinic acid 0.5 1 NA 2 64 64 114 3-ClC₆H₄ 3-ClC₆H₄3-hydroxypicolinic acid 0.125 8 NA 64 64 16 115 4-EtO-3-ClC₆H₃4-EtO-3-ClC₆H₃ 3-hydroxypicolinic acid 2 2 1 8 16 64 116 2-Cl-5-Br-6-2-F-4-ClC₆H₃ 3-hydroxypicolinic acid 2 1 0.25 4 4 64 FC₆H₂ 1172-Me-4-ClC₆H₃ 3-ClC₆H₄ 3-hydroxypicolinic acid 2 1 0.5 4 4 16 1182-Me-4-ClC₆H₃ 2-Me-4-ClC₆H₃ 3-hydroxypicolinic acid 1 0.25 0.12 1 1 16119 4-Me-3-ClC₆H₃ 4-Me-3-ClC₆H₃ 3-OAc-picolinic acid 2 1 NA 2 2 64 1204-Me-3-ClC₆H₃ 4-Me-3-ClC₆H₃ 4-hydroxybenzimidazole 0.125 4 NA 8 8 1213-ClC₆H₄ 3-ClC₆H₄ 4-hydroxyethylimidazole 0.125 4 8 32 32 4 122 3-ClC₆H₄3-ClC₆H₄ 6-amino-3-hydroxypicolinic 0.25 4 16 32 32 8 acid 123 3-ClC₆H₄3-ClC₆H₄ Imidazole acetic acid 0.125 2 8 32 32 8 Ceftriaxone 2 1 <0.12564 64 0.12 Ciprofloxacin 0.12 0.12 0.5 0.5 64 0.12 Cloxacillin 0.12 0.250.12 16 64 8 Erythromycin 0.5 0.5 NA 2 NA 4 Imipenem 0.12 0.12 <0.125 164 2 Meropenem 0.06 0.06 NA 2 NA 0.06 Pen G 0.5 16 <0.125 1 32 0.12

TABLE 3 Anti-mycobacterium In vitro Activity M. tuberculosis MIC(mcg/mL) Compound H37Rv* P2SP1** P1SP2** 10 0.387 0.387 0.387 50 0.3870.387 0.387 51 0.387 0.387 0.387 53 0.775 0.775 0.387 55 0.775 0.7750.387 65 0.775 0.775 0.775 72 0.775 0.775 0.775 75 0.775 0.775 0.775Isoniazid (INH) <0.062 >8 >8 Rifampicin <0.125 16 >16 Ethambutol <1 8 8Ethionamide 1 >64 32 p-aminosalicylate <0.25 32 16 Ofloxacin 4 32 16Streptomycin <2 <2 <2 Kanamycin <2 <2 <2 cycloserine 8 8 8*Sensitive strain**Multi-drug resistant strain

TABLE 4 Antifungal Activity for Select Borinic Acid Complexes MIC(μg/ml) Candida Candida Aspergillus albicans glabrata CandidaMicrosporum fumigatus Trichophyton ATCC ATCC parapsilosis canis ATCCmentagrophytes Compound 90028 15126 ATCC 22019 ATCC 10214 204305 ATCC10270  53 1 >32 32 16 >32 >32 124 1 >32 32 16 >32 >32 125 1 >32 3216 >32 >32 126 1 >32 32 16 >32 >32 127 1 >32 16 16 >32 32 128 1 32 1612 >32 16 129 1 >32 32 16 >32 32 130 1 0.5 0.5 0.5 0.5 0.5 131 2 >32 >3232 >32 32 132 2 1 0.5 0.38 >32 0.25 Amphotericin B 1 0.75 0.75 0.5 1 1Fluconazole 0.25 3 2.5 16 >64 16

The present invention also encompasses the acylated prodrugs of thecompounds of the invention. Those skilled in the art will recognizevarious synthetic methodologies which may be employed to preparenon-toxic pharmaceutically acceptable addition salts and acylatedprodrugs of the inventive compounds.

Tables 1 to 4 also contain inhibitory activity for known antibiotics,shown at the end of each of the tables.

EXAMPLES

Proton NMR are recorded on Varian AS 400 and MercuryPlus 300 MHzspectrometer and chemical shifts are reported as δ (ppm) down field fromtetramethylsilane. Mass spectra are determined on Micromass Quattro IIand Applied Biosystems AP3000. Compound numbers appear in parenthesesand correspond to numbers in Tables 1 to 4.

Formation of Ethylene Glycol Boronate Ester (3, T=nothing)

General Procedure

Boronic acid was dissolved in dry THF, dry toluene or dry diethyl ether(˜10 mL/g) under nitrogen. Ethylene glycol (1 molar equivalent) wasadded to the reaction and the reaction was heated to reflux for 1 to 4hours. Reaction was cooled to room temperature and solvent was removedunder reduced pressure leaving the ethylene glycol ester as an oil or asolid. In cases where an oil was obtained or a solid that dissolved inhexane, dry hexane was added and removed under reduced pressure. Theproduct was then placed under high vacuum for several hours. In caseswhere a solid was obtained that did not dissolve in hexane, the solidwas collected by filtration and washed with cold hexane.

3-Cyanophenylboronic acid ethylene glycol ester (3a)

3-Cyanophenyl boronic acid (1 g, 6.8 mmol) was dissolved in dry THF (10mL) under nitrogen. Ethylene glycol (379 μL, 422 mg, 6.8 mmol) was addedand the reaction was heated to reflux for 4 hours then cooled to roomtemperature. THF was removed by rotary evaporator to give a white solid.Cold hexane was added and the product was collected by filtration givinga white solid (1.18 g, quant. yield). ¹H-NMR (300.058 MHz, DMSO-d6) δppm 7.92-8.01 (3H, m), 7.50-7.64 (1H, m), 4.35 (4H, s)

Thiophene 3-boronic acid ethylene glycol ester (3b)

Thiophene-3-boronic acid (1 g, 7.8 mmol) was dissolved in dry THF (10mL) under nitrogen. Ethylene glycol (435 μL, 484 mg, 7.8 mmol) was addedand the reaction was heated to reflux for 1 hour then cooled to roomtemperature. THF was removed by rotary evaporator to give a white solid.Hexane was added, dissolving the solid and removed by rotaryevaporation. The product was placed under high vacuum to yield a tansolid (1.17 g, 97%). ¹H-NMR (300.058 MHz, CDCl₃) δ ppm 7.93 (1H, s),7.3-7.4 (2H, m), 4.35 (4H, s).

3-Fluorophenylboronic acid ethylene glycol ester (3c)

A mixture of 3-fluorophenylboronic acid (7.00 g, 50.0 mmol) and ethyleneglycol (2.8 mL, 50 mmol) in toluene (200 mL) was heated to reflux for 3hours under Dean-Stark conditions. The solvent was removed under reducedpressure to afford 3-fluorophenylboronic acid ethylene glycol ester(7.57 g, 91%).

Formation of Unsymmetrical Borinic Acid (6) from Boronic Acid EthyleneGlycol Ester

General Procedure A: Grignard Methodology

Boronic acid ethylene glycol ester was dissolved in dry THF (10-20 mL/g)under nitrogen. Solution was cooled to −78° C. in an acetone/dry icebath or to 0° C. in an ice/water bath. Grignard reagent (0.95 to 1.2molar equivalent) was added dropwise to the cooled solution. Thereaction was warmed to room temperature and stirred for 3-18 hours. 6NHCl (2 mL/g) was added and solvent was removed under reduced vacuum.Product was extracted into diethyl ether (40 mL/g) and washed with water(3×equal volume). Organic layer was dried (MgSO₄), filtered and thesolvent was removed by rotary evaporation giving the crude product,which is either purified by column chromatography or taken onto the nextstep without purification. Alternative work-up: if the borinic acidproduct contained a basic group such as an amine or pyridine, then afterstirring at room temperature for 3-18 hours, water (2 mL/g) was addedand the pH adjusted to 5-8. Product was extracted into diethyl ether orethyl acetate or THF up to three times (40 mL/g). Organic layer wasdried (MgSO₄), filtered and the solvent was removed by rotaryevaporation giving the crude product, which is either purified by columnchromatography or taken onto the next step without purification.

(4-Cyanophenyl)(3-fluorophenyl)borinic acid (6a)

4-Cyanophenyl boronic acid ethylene glycol ester (500 mg, 2.89 mmol) wasdissolved in dry THF under nitrogen. The solution was cooled to −78° C.in an acetone/dry ice bath and 3-fluorophenylmagnesium bromide (1M inTHF)(2.74 mL, 2.74 mmol, 0.95 molar equivalent) was added dropwise tothe cold solution. The reaction was allowed to warm slowly to roomtemperature and stirred for 18 hours. 6N HCl (1 mL) was added to thereaction causing a cloudy appearance and the solvent was removed using arotary evaporator. The product was extracted into diethyl ether (20 mL)and washed with water (3×20 mL). The organic layer was dried (MgSO4),filtered and the solvent removed using a rotary evaporator to yield thecrude product as an oily solid. This was taken onto the next stepwithout purification.

General Procedure B: (Hetero)aryl-lithium Methodology

The (hetero)aryl-bromide or iodide was dissolved in dry THF (20-30 mL/g)under nitrogen and degassed. The solution was cooled to −78° C. in anacetone/dry ice bath and n-, sec- or tert-butyllithium in THF or othersolvent (1.2-2.4 molar equivalents) was added to the cooled solutiondropwise (generally causing the solution to turn deep yellow). Theboronic acid ethylene glycol ester (1 molar equivalent) was dissolved indry THF or diethyl ether (2-10 mL/g) under nitrogen. The boronic acidethylene glycol ester in THF was added dropwise to the cooledaryl-lithium solution (generally causing the solution to turn paleyellow). The reaction was warmed to room temperature and stirred for1-18 hours. 6N HCl (2-4 mL/g) was added and solvent was removed underreduced vacuum. Product was extracted into diethyl ether (40 mL/g) andwashed with water (3×equal volume). Organic layer was dried (MgSO₄),filtered and the solvent was removed by rotary evaporation giving thecrude product, which is either purified by column chromatography ortaken onto the next step without purification. Alternative work-up: ifthe borinic acid product contained a basic group such as an amine orpyridine then after stirring at room temperature for 3-18 hours water (2mL/g) was added and the pH adjusted to 5-8. Product was extracted intodiethyl ether or ethyl acetate or THF up to three times (40 mL/g) andwashed with water (3×equal volume). Organic layer was dried (MgSO₄),filtered and the solvent was removed by rotary evaporation giving thecrude product, which is either purified by column chromatography ortaken onto the next step without purification.

(3-Thienyl)(3-chlorophenyl)borinic acid (6b)

3-Chloro-bromobenzene (447 μL, 728 mg, 3.8 mmol) was dissolved in dryTHF (15 mL) under nitrogen. The solution was degassed and cooled to −78°C. in an acetone/dry ice bath. tert-Butyllithium (1.7M in THF)(4.47 mL,7.6 mmol, 2 molar equivalent) was added to the cooled solution dropwisecausing the solution to turn deep yellow. The solution was stirred at−78° C. while 3-thiopheneboronic acid ethylene glycol ester (586 mg) wasdissolved in dry diethyl ether (1 mL). The boronic ester solution wasthen added dropwise to the cooled solution causing the color to changeto pale yellow. The reaction was warmed to room temperature and stirredfor 18 hours. 6N HCl (2 mL) was added and the reaction was stirred for 1hour. The solvent was removed using a rotary evaporator. The product wasextracted into diethyl ether (10 mL) and washed with water (2×10 mL).The organic layer was dried (MgSO4), filtered and the solvent removedusing a rotary evaporator to yield the crude product as an orange oil.The product was purified by column chromatography using silica gel andhexane: ethyl acetate 5:1 as eluent giving the pure product as a clearoil (614 mg, 73%).

(3-Chlorophenyl)vinylborinic acid (6c)

This was prepared by a similar process as described for 6b by thereaction of 3-cyanophenyl boronic acid ethylene glycol ester withvinyllithium.

(3-Fluoro-5-chlorophenyl)ethynylborinic acid (6d)

This was prepared by a similar process as described for 6b by thereaction of 3-fluoro-5-chlorophenyl boronic acid ethylene glycol esterwith ethynyllithium.

(4-Methyl-3-chlorophenyl)(2-thienyl)borinic acid (6e)

This was prepared by a similar process as described for 6b by thereaction of 2-thienylboronic acid ethylene glycol ester with4-methyl-3-chlorophenyllithium.

(4-Cyanophenyl)ethynylborinic acid (6f)

This was prepared by a similar process as described for 6b by thereaction of 4-cyanophenylboronic acid ethylene glycol ester withethynyllithium.

(3-Fluorophenyl)cyclopropylborinic acid (6q)

This was prepared by a similar process as described for 6b by thereaction of 3-fluorophenylboronic acid ethylene glycol ester withcyclopropyllithium.

(3-Thienyl)methylborinic acid (6h)

This was prepared by a similar process as described for 6b by thereaction of 3-thienylboronic acid ethylene glycol ester withmethyllithium.

(4-Pyridyl)phenylborinic acid (6i)

This was prepared by a similar process as described for 6b by thereaction of phenylboronic acid ethylene glycol ester with4-pyridyllithium.

(3-Cyanophenyl)(2-fluorophenyl)borinic acid (6j)

This was prepared by a similar process as described for 6b by thereaction of 3-cyanophenylboronic acid ethylene glycol ester with2-fluorophenyllithium.

4-(Dimethylaminomethyl)phenyl 3-fluorophenyl borinic acid (6k)

Sec-butyllithium (1.4 M in cyclohexane, 6.0 mL) was added to a solutionof N,N-dimethyl-4-bromobenzylamine (1.50 g, 7.00 mmol) in THF (14 mL) at−78° C. under nitrogen atmosphere and the mixture was stirred for 15min. 3-Fluorophenylboronic acid ethylene glycol ester (1.16 g, 7.00mmol) in THF (7 mL) was added to the mixture. The reaction was allowedto warm to room temperature and stirred for 1 h. Water was added and themixture was washed with ether. The pH was adjusted to 8 with 1Mhydrochloric acid. The mixture was extracted with ethyl acetate twice.The organic layer was washed with brine and dried on anhydrous sodiumsulfate. The solvent was removed under reduced pressure to afford theborinic acid (890 mg, 49%).

Formation of Symmetrical Borinic Acid (5) by Reaction of Organometallicswith Trialkyl Borates.

Bis(4-chlorophenyl)borinic acid (5a) (Procedure C)

A cold solution (−78° C.) of trimethyl borate (0.37 ml) in drytetrahydrofuran (THF, 25 ml) was treated dropwise with4-chlorophenylmagnesium bromide (6.75 ml, 1M solution in ether). Thereaction mixture was stirred at −78° C. for 1 h and then stirred for 18h at room temperature. The solvent was removed under reduced pressure.The resultant residue was stirred with 100 ml of ether and 15 ml of 6Nhydrochloric acid. Organic layer was separated and aqueous layer wasextracted with ether (2×100 ml). The combined organic extract was washedwith brine and dried over anhydrous magnesium sulfate. Solvent wasremoved to give light yellowish solid. The product was chromatographedover silica gel (Hex:Ether=1:1) to give 420 mg of borinic acid. ¹H NMR(400 MHz, CDCl₃) δ: 5.84 (s, OH), 7.46 (d, 4H, Ar—H), 7.72 (d, 4H,Ar—H).

Bis(3-Chloro-4-methylphenyl)borinic acid (5b)

In a similar manner as for 5a, the titled compound was obtained from thereaction of 3-chloro4-methylphenylmagnesium bromide with trimethylborate. The product was obtained by chromatography over silica gel.

Bis(3-Fluoro-4-methylphenyl)borinic acid (5c)

In a similar manner as for 5a, the titled compound was obtained from thereaction of 3-fluoro-4-methylphenyllithium with trimethyl borate. Theproduct was obtained by chromatography over silica gel.

Bis(3-Chloro-4-methoxyphenyl)borinic acid (5d)

In a similar manner as for 5a, the titled compound was obtained from thereaction of 3-chloro-4-methoxyphenyllithium with trimethyl borate. Theproduct was obtained by chromatography over silica gel.

Bis(3-Fluoro-4-methoxyphenyl)borinic acid (5e)

In a similar manner as for 5a, the titled compound was obtained from thereaction of 3-fluoro-4-methoxyphenyllithium with trimethyl borate. Theproduct was obtained by chromatography over silica gel.

Formation of Unsymmetrical Borinic Acids (6) by Reaction ofOrganometallics with alkyl(or aryl or alkenyl)dialkoxyboranes.

(4-Chloro-phenyl)methyl-borinic acid (6m) (Procedure D)

To 4-chlorophenylmagnesium bromide (5.5 ml, 1M solution in ether) at−78° C., di(isopropoxy)methylborane (1 ml, 0.78 g) was added dropwisevia syringe. The reaction mixture was stirred at −78° C. for 1 h andthen stirred overnight at ambient temperature. The reaction mixture wastreated dropwise with 100 ml of ether and 15 ml of 6N hydrochloric acid,and stirred for 1 h. Organic layer was separated and aqueous layer wasextracted with ether (2×100 ml). The combined organic extract was washedwith brine and dried over anhydrous sodium sulfate. Solvent was removedunder reduce pressure to give 1.1 g of oil. ¹H NMR of the product wasconsistent for (4-chlorophenyl)methyl borinic acid.

(4-Fluorophenyl)methylborinic acid (6n)

In a similar manner as for 6m, the titled compound was obtained from thereaction of 4-fluorophenylmagnesium bromide withdi(isopropoxy)methylborane. The product was obtained by chromatographyover silica gel.

(4-Biphenyl)methylborinic acid (6o)

In a similar manner as for 6m, the titled compound was obtained from thereaction of 4-biphenyllithium with di(isopropoxy)methylborane. Theproduct was obtained by chromatography over silica gel.

(3-Chloro-4-methylphenyl)methylborinic acid (6p)

In a similar manner as for 6m, the titled compound was obtained from thereaction of 3-chloro4-methylphenyllithium withdi(isopropoxy)methylborane. The product was obtained by chromatographyover silica gel.

(3-Chloro-4-methoxyphenyl)methylborinic acid (6q)

In a similar manner as for 6m, the titled compound was obtained from thereaction of 3-chloro4-methoxyphenyllithium withdi(isopropoxy)methylborane. The product was obtained by chromatographyover silica gel.

(4-Dimethylaminophenyl)methylborinic acid (6r)

In a similar manner as for 6m, the titled compound was obtained from thereaction of 4-dimethylaminophenyllithium withdi(isopropoxy)methylborane. The product was obtained by chromatographyover silica gel.

(3-Pyridyl)vinyl borinic acid (6s)

Isopropylmagnesium chloride (2.0 M in THF) (5.0 mL, 10 mmol) was addedto a solution of 3-bromopyridine (1.60 g, 10.0 mmol) in THF (15 mL)under nitrogen atmosphere at room temperature and the mixture wasstirred for 1 h. Vinylboronic acid dibutyl ester (3.4 mL) was added tothe reaction dropwise and the mixture was stirred at room temperaturefor 18 h. Water was added and the pH was adjusted to 7 with 1 Mhydrochloric acid. The mixture was extracted with ethyl acetate. Theorganic layer was washed with brine and dried on anhydrous sodiumsulfate. The solvent was removed under reduced pressure to give thetitle compound (1.04 g, 78%).

(3-Chloro-4-dimethylaminophenyl)vinylborinic acid (6t)

In a similar manner as for 6s, the titled compound was obtained from thereaction of 3-chloro4-dimethylaminophenyllithium with vinylboronic aciddibutyl ester. The product was obtained by chromatography over silicagel.

Borinic Acid-Alkylalcohol Derivatives

Bis(3-Chlorophenyl)borinic acid 4-(hydroxyethyl)imidazole ester (121)

To a solution of bis(3-chlorophenyl)borinic acid (0.4 g, 1.428 mmol) inethanol (10 ml), 4-(hydroxyethyl)imidazole hydrochloride (0.191 g, 1.428mmol), sodium bicarbonate (0.180 g, 2.143 mmol) were added and thereaction mixture was stirred at room temperature for 18 h. Salt wasremoved by filtration. Filtrate was concentrated and treated with hexaneto afford the product as a solid and was collected by filtration. (450mg, 84.9% yield). ¹H NMR (CD₃OD) δ (ppm) 2.92 (t, 2H), 3.82 (t, 2H),7.0-7.2 (m, 9H), 7.90 (s, 1 H); (ES⁻)(m/z) 343.11, MF C₁₇H₁₅BCl₂N₂O

Bis(4-Chlorophenyl)borinic acid 4-(hydroxymethyl)imidazole ester (126)

In a similar manner as in Example 121, the titled compound was obtainedfrom the reaction of bis(4-chlorophenyl)borinic acid with4-(hydroxymethyl)imidazole hydrochloride. The product was obtained aswhite crystals. (ES⁻)(m/z) 328.79, MF C₁₆H₁₃BCl₂N₂O

Bis(3-Chloro-4-methylphenyl)borinic acid1-benzyl4-(hydroxymethyl)-imidazole ester (127)

To a solution of 1-benzyl-4-(hydroxymethyl)imidazole (96 mg, 0.521 mmol)in methanol (5 ml), bis(3-chloro4-methylphenyl)borinic acid (121 mg,0.521 mmol) was added and the reaction mixture was stirred at roomtemperature for 2 h. Solvent was removed under reduced pressure and theresidue was treated with hexane to give a solid. The product wasisolated by filtration and washed with hexane to give product (193 mg,83%). ¹H NMR (CDCl₃) δ: 2.3 (s, 6H, 2×CH₃), 4.8 (brs, 2H, CH₂), 5.1(brs, 2H, CH₂), 6.9-7.4 (complex, 13H, Ar—H); MS (ES⁺)(m/z) 448.78, MFC₂₅H₂₃BCl₂N₂O.

Bis(3-Chloro-4-methylphenyl)borinic acid1-methyl-2-(hydroxymethyl)-imidazole ester (128)

In a similar manner as in Example 127, the titled compound was obtainedfrom the reaction of bis(3-chloro4-methylphenyl)borinic acid with1-methyl-2-(hydroxy-methyl)imidazole hydrochloride. The product wasobtained as white crystals. (ES⁺)(m/z) 372.82, MF C₁₉H₂₁BCl₂N₂O

Bis(3-Chloro-4-methylphenyl)borinic acid1-ethyl-2-(hydroxymethyl)-imidazole ester (129)

In a similar manner as in Example 127, the titled compound was obtainedfrom the reaction of bis(3-chloro-4-methylphenyl)borinic acid with1-ethyl-2-(hydroxy-methyl)imidazole hydrochloride. The product wasobtained as white crystals. (ES⁺)(m/z) 386.83, MF C₂₀H₂₃BCl₂N₂O

Bis(3-Chloro-4-methylphenyl)borinic acid1-methyl4-(hydroxymethyl)-imidazole ester (130)

In a similar manner as in Example 127, the titled compound was obtainedfrom the reaction of bis(3-chloro-4-methylphenyl)borinic acid with1-methyl-4-(hydroxy-methyl)imidazole hydrochloride. The product wasobtained as white crystals. (ES⁺)(m/z) 372.88, MF C₁₉H₂₁BCl₂N₂O

Bis(3-Chloro-4-methylphenyl)borinic acid 2-pyridylethanol (131)

In a similar manner as in Example 121, the titled compound was obtainedfrom the reaction of bis(3-chloro-4-methylphenyl)borinic acid with2-pyridylethanol. The product was obtained as white crystals. (ES⁺)(m/z)383.84, MF C₂₁H₂₀BCl₂NO

Hydroxyquinoline Derivatives

Bis(3-Chlorophenyl)borinic acid 5-cyano8-hydroxyquinoline ester (19)

To a solution of bis(3-chlorophenyl)borinic acid (0.25 g) in ethanol (5ml) and water (2 ml) was added 5-cyano-8-hydroxyquinoline (0.15 g). Thesolution was stirred at room temperature for 21 hours. A yellow solidprecipitate formed which was collected by filtration and washed withcold ethanol. The product was obtained as yellow crystals. ¹H NMR(DMSO-d6) δ (ppm) 7.24-7.35 (m, 8H),7.38 (d, 1H), 8.18 (dd, 1H), 8.40(d, 1H), 8.86 (d, 1H), 9.50 (d, 1H).

(3-Chlorophenyl)(2-thienyl)borinic acid 8-hydroxyquinoline ester (36)

To a solution of (3-chlorophenyl)(2-thienyl)borinic acid (1.5 g) inethanol (2 ml) was added 8-hydroxyquinoline (0.77 g) in hot ethanol (2ml). The reaction was heated to reflux and cooled to room temperature. Ayellow solid precipitated. The mixture was cooled in ice, the solid wascollected by filtration and washed with cold ethanol. The product wasobtained as a yellow solid (1.01 g). ¹H NMR (DMSO) δ (ppm) 6.98-7.06 (m,2H), 7.19-7.26 (m, 3H), 7.38-7.50 (m, 4H), 7.71 (t, 1H), 7.91 (dd, 1H),8.80 (d, 1H), 9.18 (d, 1H); (ESI⁺)(m/z) 350.1, MF C₁₉H₁₃BClNOS

(2-Thienyl)methylborinic acid 8-hydroxyquinoline ester (26)

In a similar manner as in Example 36, the titled compound was obtainedfrom the reaction of (2-thienyl)methylborinic acid with8-hydroxyquinoline. The product was obtained as yellow crystals.

(3-Cyanophenyl)vinylborinic acid 8-hydroxyquinoline ester (40)

In a similar manner as in Example 36, the titled compound was obtainedfrom the reaction of (3-cyanophenyl)vinylborinic acid with8-hydroxyquinoline. The product was obtained as yellow crystals.(ESI⁺)(m/z) 285.1, MF C₁₈H₁₃BN₂O

(2-Chlorophenyl)ethynylborinic acid 8-Hydroxyquinoline ester (43)

In a similar manner as in Example 36, the titled compound was obtainedfrom the reaction of (2-chlorophenyl)ethynylborinic acid with8-hydroxyquinoline. The product was obtained as yellow crystals.(ESI⁺)(m/z) 292.1, MF C₁₇H₁₁BClNO

Bis(ethynyl)borinic acid 8-Hydroxyquinoline (44)

In a similar manner as in Example 36, the titled compound was obtainedfrom the reaction of bis(ethynyl)borinic acid with 8-hydroxyquinoline.The product was obtained as light yellow crystals. (ESI⁺)(m/z) 206.1, MFC₁₃H₈BNO

(3-Fluorophenyl)cyclopropylborinic acid 8-hydroxyquinoline ester (70)

In a similar manner as in Example 36, the titled compound was obtainedfrom the reaction of (3-fluorophenyl)cyclopropylborinic acid with8-hydroxyquinoline. The product was obtained as light yellow crystals.(ES⁻)(m/z) 291.05, MF C₁₈H₁₅BFNO

(3-Pyridyl)vinylborinic acid 8-hydroxyquinoline ester (99)

A solution of (3-pyridyl)vinyl borinic acid (1.04 g, 7.82 mmol) and8-hydroxyquinoline (961 mg, 6.63 mmol) in ethanol was stirred at 40° C.for 20 min. The solvent was removed under reduced pressure and theresidue was crystallized from diethyl ether/diisopropyl ether/hexane toafford the title product (99) as a light yellow powder (355 mg, 21%). ¹HNMR (DMSO-d₆) δ (ppm) 5.23 (dd, 1H), 5.46 (dd, 1H), 6.43 (dd, 1H), 7.14(d, 1H), 7.19 (dd, 1H), 7.41 (d, 1H), 7.6-7.8 (m, 2H), 7.88 (dd, 1H),8.35 (dd, 1H), 8.57 (s, 1H), 8.76 (d, 1H), 9.00 (d, 1H); ESI⁺ (m/z) 261MF C₁₆H₁₃BN₂O.

(4-(Dimethylaminomethyl)phenyl)(3-fluorophenyl)borinic acid8-hydroxy-quinoline ester (100)

In a similar manner as in Example 99, the titled compound was obtainedfrom the reaction of(4-(Dimethylaminomethyl)phenyl)(3-fluorophenyl)borinic acid with8-hydroxyquinoline. The product was obtained as a light yellow powder.ESI⁺ (m/z) 385 MF C₂₄H₂₂BFN₂O.

3-Hydroxypicolinic Acid Derivatives

Bis(3-Chloro4-methylphenyl)borinic acid 3-hydroxypicolinate ester (111)

Bis(3-chloro4-methylphenyl)borinic acid (14.6 g) was dissolved inethanol (120 ml) and heated to reflux. 3-Hydroxypicolinic acid (5.83 g)was added in portions to the hot solution. The reaction was stirred atreflux for 15 minutes after the addition of the last portion of3-hydroxypicolinic acid was added and then cooled to room temperature.Reaction was concentrated by removal of some ethanol. Solid was removedby filtration. One recrystallization from ethanol afforded the titleproduct as white crystals (13.4 g). MP=165.0-166.5° C.

In a preferred embodiment, the present invention includes the compoundsspecifically recited herein, and pharmaceutically acceptable saltsthereof, and compositions of any of these compounds where these comprisea pharmaceutically acceptable carrier.

The present invention also relates to a method for treating amicrobial-caused disease in a patient afflicted therewith and/orpreventing such infection in a patient at risk of becoming so-infected,comprising administering to said patient a therapeutically effectiveamount of any of the compounds of the invention, preferably one or moreof those listed in Tables 1 to 4. In one aspect, the compounds of theinvention have anti-bacterial (i.e., bactericidal) and anti-fungal(i.e., fungicidal) activity.

In a preferred embodiment, the microbe is a bacterium, preferably a grampositive bacterium, wherein said gram positive bacterium is a memberselected from the group consisting of Staphylococcus species,Streptococcus species, Bacillus species, Mycobacterium species,Corynebacterium species, Clostridium species, Actinomyces species,Enterococcus species, and Streptomyces species.

In another preferred embodiment of such method, the bacterium is a gramnegative bacterium, preferably one selected from the group consisting ofAcinetobacter species, Neisseria species, Pseudomonas species, Brucellaspecies, Agrobacterium species, Bordetella species, Escherichia species,Shigella species, Yersinia species, Salmonella species, Klebsiellaspecies, Enterobacter species, Haemophilus species, Pasteurella species,Streptobacillus species, spirochetal species, Campylobacter species,Vibrio species, and Helicobacter species.

In a highly preferred embodiment of the present invention, the bacteriumis a member selected from the group consisting of Staphylococcus aureus;Staphylococcus epidermidis; Staphylococcus saprophyticus; Streptococcuspyogenes; Streptococcus agalactiae; Streptococcus pneumoniae;Enterococcus faecalis; Enterococcus faecium; Bacillus anthracis;Mycobacterium avium; Mycobacterium tuberculosis; Acinetobacter baumanii;Corynebacterium diphtheria; Clostridium perfringens; Clostridiumbotulinum; Clostridium tetani; Neisseria gonorrhoeae; Neisseriameningitidis; Pseudomonas aeruginosa; Legionella pneumophila;Escherichia coli; Yersinia pestis; Haemophilus influenzae; Helicobacterpylori; Campylobacter fetus; Campylobacter jejuni; Vibrio cholerae;Vibrio parahemolyticus; Trepomena pallidum; Actinomyces israelii;Rickettsia prowazekii; Rickettsia rickettsii; Chlamydia trachomatis;Chlamydia psittaci; Brucella abortus; Agrobacterium tumefaciens; andFrancisella tularensis.

In a preferred embodiment the microbe is a fungus or yeast wherein saidfungus is a member selected from the group consisting of Aspergillusspecies, Trichophyton species, Microsporium species, Cryptococcusneoformans, Blastomyces dermatitidis, Coccidiodes immitis, Histoplasmacapsulatum, or Paracoccidioides brasiliensis and wherein said yeast is amember selected from the group consisting of Candida albicans, Candidaglabrata, Candida krusei, Candida tropicalis, or Candida parapsilosis.

1. A compound having the structure of Formula 1

wherein B is boron, O is oxygen wherein R* and R** are eachindependently selected from substituted or unsubstituted alkyl (C₁- C₄),substituted or unsubstituted cycloalkyl (C₃- C₇), substituted orunsubstituted alkenyl, substituted or unsubstituted alkynyl, substitutedor unsubstituted aralkyl, substituted or unsubstituted phenyl, andsubstituted or unsubstituted heteroaryl, and wherein z is 0 or 1 andwhen z is 1, A is CH, CR¹⁰ or N, and wherein D is N, CH, or CR¹²,wherein R¹² is selected from (CH₂)_(k)OH (where k=1, 2 or 3), CH₂NH₂,CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H, CO₂alkyl, CONH₂, OH, alkoxy, aryloxy,SH, S-alkyl, S-aryl, SO₂N(alkyl)₂, SO₂NHalkyl, SO₂NH₂, SO₂alkyl, SO₃H,SCF₃, CN, halogen, CF₃, NO₂, NH₂, 2*-amino, 3*-amino, NH₂SO₂ and CONH₂,and wherein J is CR¹⁰ or N and wherein R⁹, R¹⁰ and R¹¹ are eachindependently selected from the group consisting of hydrogen, alkyl,cycloalkyl, (CH₂)_(n)OH (n=2 to 3), CH₂NH₂, CH₂NHalkyl, CH₂N(alkyl)₂,halogen, CHO, CH═NOH, CO₂H, CO₂-alkyl, S-alkyl, SO₂-alkyl, S-aryl,SO₂N(alkyl)₂, SO₂NHalkyl, SO₂NH₂, NH₂, alkoxy, CF₃, SCF₃, NO₂, SO₃H andOH, including salts thereof.
 2. The compound of claim 1 wherein R* andR** are the same.
 3. The compound of claim 1 wherein R* and R** aredifferent.
 4. The compound of claim 1 wherein one of R* and R** is asubstituted or unsubstituted alkyl (C₁-C₄).
 5. The compound of claim 1wherein R* and R** are each a substituted or unsubstituted alkyl(C₁-C₄).
 6. The compound of claim 1 wherein one of R* and R** is asubstituted or unsubstituted cycloalkyl (C₃- C₇).
 7. The compound ofclaim 1 wherein R* and R** are each a substituted or unsubstitutedcycloalkyl (C₃-C₇).
 8. The compound of claim 1 wherein one of R* and R**is a substituted or unsubstituted alkenyl.
 9. The compound of claim 1wherein R* and R** are each a substituted or unsubstituted alkenyl. 10.The compound of claim 8 or 9 wherein said alkenyl has the structure

wherein R¹, R², and R³ are each independently selected from the groupconsisting of hydrogen, alkyl, aryl, cycloalkyl, substituted aryl,aralkyl, substituted aralkyl, (CH₂)_(k)OH (where k=1, 2 or 3), CH₂NH₂,CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H, CO₂alkyl, CONH₂, S-alkyl, S-aryl,SO₂alkyl, SO₂N(alkyl)₂, SO₂NHalkyl, SO₂NH₂, SO₃H, SCF₃, CN, halogen, CF₃and NO₂.
 11. The compound of claim 1 wherein one of R* and R** is asubstituted or unsubstituted alkynyl.
 12. The compound of claim 1wherein R* and R** are each a substituted or unsubstituted alkynyl. 13.The compound of claim 11 or 12 wherein said alkynyl has the structure

wherein R¹ is selected from the group consisting of hydrogen, alkyl,cycloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl,(CH₂)_(k)OH (where k=1, 2 or 3), CH₂NH₂, CH₂NH-alkyl, CH₂N(alkyl)₂,CO₂H, CO₂alkyl, CONH₂, S-alkyl, S-aryl, SO₂alkyl, SO₂N(alkyl)₂,SO₂NHalkyl, SO₂NH₂, SO₃H, SCF₃, CN, halogen, CF₃ and NO₂.
 14. Thecompound of claim 1 wherein one of R* and R** is a substituted orunsubstituted phenyl.
 15. The compound of claim 1 wherein R* and R** areeach a substituted or unsubstituted phenyl and wherein when J is —CH—and z=1, then A and D are not both —CH—.
 16. The compound of claim 12 or13 wherein said phenyl has the structure

wherein R⁴, R⁵, R⁶, R⁷ and R⁸ are each independently selected from thegroup consisting of hydrogen, alkyl, cycloalkyl aryl, substituted aryl,aralkyl, substituted aralkyl, (CH₂)_(k)OH (where k=1, 2 or 3), CH₂NH₂,CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H, CO₂alkyl, CONH₂, CONHalkyl,CON(alkyl)₂, OH, alkoxy, aryloxy, SH, S-alkyl, S-aryl, SO₂alkyl,SO₂N(alkyl)₂, SO₂NHalkyl, SO₂NH₂, SO₃H, SCF₃, CN, halogen, CF₃, NO₂,NH₂, 2^(o)-amino, 3^(o)-amino, NH₂SO₂, OCH₂CH₂NH₂, OCH₂CH₂NHalkyl,OCH₂CH₂N(alkyl)₂, oxazolidin-2-yl, or alkyl substituted oxazolidin-2-yl.17. The compound of claim 1 wherein R* is 3-fluorophenyl, R** is4-chlorophenyl, R⁹ is H, R¹¹ is H, A_(z) is CH, D is CH, J is CH. 18.The compound of claim 1 wherein R* and R** are each3-(4,4-dimethyloxazolidin-2-yl)phenyl, and R⁹ is H, R¹¹ is H, z=1, A isCH, D is CH, J is CH.
 19. The compound of claim 1 wherein R* is3-fluorophenyl, R** is cyclopropyl, R⁹ is H, R¹¹ is H, z=1, A is CH, Dis CH, and J is CH.
 20. The compound of claim 1 wherein R* is4-(N,N-dimethyl)-aminomethylphenyl, R** is 4-cyanophenyl, R⁹ is H, R¹¹is H, z=1, A is CH, D is CH, and J is CH.
 21. The compound of claim 1wherein one of R* and R** is a substituted or unsubstituted benzyl. 22.The compound of claim 1 wherein R* and R** are each a substituted orunsubstituted benzyl.
 23. The compound of claim 21 or 22 wherein saidbenzyl has the structure

wherein R⁴, R⁵, R⁶, R⁷ and R⁸ are each independently selected from thegroup consisting of alkyl, cycloalkyl, aryl, substituted aryl, aralkyl,substituted aralkyl, (CH₂)_(k)OH (where k=1, 2 or 3), CH₂NH₂,CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H, CO₂alkyl, CONH₂, CONHalkyl,CON(alkyl)₂, OH, alkoxy, aryloxy, SH, S-alkyl, S-aryl, SO₂alkyl,SO₂N(alkyl)₂, SO₂NHalkyl, SO₂NH₂, SO₃H, SCF₃, CN, halogen, CF₃, NO₂,NH₂, 2^(o)-amino, 3^(o)-amino, NH₂SO₂, OCH₂CH₂NH₂, OCH₂CH₂NHalkyl,OCH₂CH₂N(alkyl)₂, oxazolidin-2-yl, or alkyl substituted oxazolidin-2-yl.24. The compound of claim 1 wherein one of R* and R** is a substitutedor unsubstituted heterocycle.
 25. The compound of claim 1 wherein R* andR** are each a substituted or unsubstituted heterocycle.
 26. Thecompound of claim 18 or 19 wherein said heterocycle has the structure

wherein X═CH═CH, N═CH, NR¹³ (wherein R¹³═H, alkyl, aryl or aralkyl), O,or S and wherein Y═CH or N and wherein R¹, R², and R³ are eachindependently selected from the group consisting of hydrogen, alkyl,cycloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl,(CH₂)_(k)OH (where k=1, 2 or 3), CH₂NH₂, CH₂NH-alkyl, CH₂N(alkyl)₂,CO₂H, CO₂alkyl, CONH₂, S-alkyl, S-aryl, SO₂alkyl, SO₂N(alkyl)₂,SO₂NHalkyl, SO₂NH₂, SO₃H, SCF₃, CN, halogen, CF₃, NO₂, oxazolidin-2-yl,or alkyl substituted oxazolidin-2-yl.
 27. The compound of claim 1wherein R* is pyrid-3-yl, R** is 4-chlorophenyl, R⁹ is H, R¹¹ is H, z=1,A is CH, D is CH, and J is CH.
 28. The compound of claim 1 wherein R* is5-cyanopyrid-3-yl, R** is vinyl, R⁹ is H, R¹¹ is H, z=1, A is CH, D isCH, and J is CH.
 29. The compound of claim 1 wherein R⁹ is H, R¹¹ is H,z=1, A is CH, D is CH, and J is CH
 30. The compound of claim 1 furthercomprising a solvent adduct having the structure

and wherein R*** is H or alkyl.
 31. A compound having the structure ofFormula 2

wherein B is boron, O is oxygen wherein R* and R** are eachindependently selected from substituted or unsubstituted alkyl (C₁-C₄),substituted or unsubstituted cycloalkyl (C₃-C₇), substituted orunsubstituted alkenyl, substituted or unsubstituted alkynyl, substitutedor unsubstituted aralkyl, substituted or unsubstituted phenyl, andsubstituted or unsubstituted heterocycle, and wherein z is 0 or 1 andwhen z is 1, A is CH, CR¹⁰ or N, and wherein D is N, CH, or CR¹², andwherein E is H, OH, alkoxy or 2-(morpholino)ethoxy, CO₂H or CO₂alkyl andwherein m=0-2, and wherein r is 1 or 2, and wherein when r is 1, G is ═O(double-bonded oxygen) and when r is 2, each G is independently H,methyl, ethyl or propyl, wherein R¹² is selected from (CH₂)_(k)OH (wherek=1, 2 or 3), CH₂NH₂, CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H, CO₂alkyl, CONH₂,OH, alkoxy, aryloxy, SH, S-alkyl, S-aryl, SO₂alkyl, SO₂N(alkyl)₂,SO₂NHalkyl, SO₂NH₂, SO₃H, SCF₃, CN, halogen, CF₃, NO₂, NH₂, 2*-amino,3*-amino, NH₂SO₂ and CONH₂, and wherein R⁹ is selected from the groupconsisting of hydrogen, alkyl, cycloalkyl, (CH₂)_(n)OH (n=1 to 3),CH₂NH₂, CH₂NHalkyl, CH₂N(alkyl)₂, halogen, CHO, CH═NOH, CO₂H, CO₂-alkyl,S-alkyl, SO₂-alkyl, S-aryl, SO₂N(alkyl)₂, SO₂NHalkyl, SO₂NH₂, NH₂,alkoxy, CF₃, SCF₃, NO₂, SO₃H and OH, including salts thereof.
 32. Thecompound of claim 31 wherein R* and R** are the same.
 33. The compoundof claim 31 wherein R* and R** are different.
 34. The compound of claim31 wherein one of R* and R** is a substituted or unsubstituted alkyl(C₁-C₄).
 35. The compound of claim 31 wherein R* and R** are each asubstituted or unsubstituted alkyl (C₁-C₄).
 36. The compound of claim 31wherein one of R* and R** is a substituted or unsubstituted cycloalkyl(C₃-C₇).
 37. The compound of claim 31 wherein R* and R** are each asubstituted or unsubstituted cycloalkyl (C₃-C₇).
 38. The compound ofclaim 31 wherein one of R* and R** is a substituted or unsubstitutedalkenyl.
 39. The compound of claim 31 wherein R* and R** are each asubstituted or unsubstituted alkenyl.
 40. The compound of claim 38 or 39wherein said alkenyl has the structure

wherein R¹, R², and R³ are each independently selected from the groupconsisting of hydrogen, alkyl, cycloalkyl, aryl, substituted aryl,aralkyl, substituted aralkyl, (CH₂)_(k)OH (where k=1, 2 or 3), CH₂NH₂,CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H, CO₂alkyl, CONH₂, S-alkyl, S-aryl,SO₂alkyl, SO₂N(alkyl)₂, SO₂NHalkyl, SO₂NH₂, SO₃H, SCF₃, CN, halogen, CF₃and NO₂.
 41. The compound of claim 31 wherein one of R* and R** is asubstituted or unsubstituted alkynyl.
 42. The compound of claim 31wherein R* and R** are each a substituted or unsubstituted alkynyl. 43.. The compound of claim 41 or 42 wherein said alkynyl has the structure

wherein R¹ is selected from the group consisting of hydrogen, alkyl,cycloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl,(CH₂)_(k)OH (where k=1, 2 or 3), CH₂NH₂, CH₂NH-alkyl, CH₂N(alkyl)₂,CO₂H, CO₂alkyl, CONH₂, S-alkyl, S-aryl, SO₂alkyl, SO₂N(alkyl)₂,SO₂NHalkyl, SO₂NH₂, SO₃H, SCF₃, CN, halogen, CF₃ and NO₂.
 44. Thecompound of claim 31 wherein one of R* and R** is a substituted orunsubstituted phenyl.
 45. The compound of claim 31 wherein R* and R**are each other than a phenyl or substituted phenyl.
 46. The compound ofclaim 31 wherein one of R* or R** is benzyl or substituted benzyl. 47.The compound of claim 31 wherein r is 1, G is ═O, m is 0 and E is OH.48. The compound of claim 31 wherein z is 1 and R⁹ is selected fromalkyl (greater than C₄), (CH₂)_(n)OH (n=1, 2 or 3), CH₂NH₂, CH₂NHalkyl,CH₂N(alkyl)₂, CHO, CH═NOH, CO₂H, CO₂-alkyl, S-alkyl, SO₂-alkyl, S-aryl,alkoxy (greater than C₄), SCF₃, and NO₂.
 49. The compound of claim 31wherein z is 1 and R¹⁰ is selected from alkyl (greater than C₄),(CH₂)_(n)OH (n=1, 2 or 3), CH₂NH₂, CH₂NHalkyl, CH₂N(alkyl)₂, CHO,CH═NOH, CO₂H, CO₂-alkyl, S-alkyl, SO₂-alkyl, S-aryl, alkoxy (greaterthan C₄), SCF₃, and NO₂.
 50. The compound of claim 31 wherein z is 1 andD is CR¹² wherein R¹² is selected from (CH₂)_(k)OH (where k=1, 2 or 3),CH₂NH₂, CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H, CO₂alkyl, CONH₂, OH, alkoxy(greater than C₄), aryloxy, SH, S-alkyl, S-aryl, SO₂alkyl, SO₃H, SCF₃,CN, NO₂, NH₂SO₂ and CONH₂.
 51. The compound of claim 31 wherein z is 1,E is N-(morpholinyl)ethoxy or alkoxy greater than C₄.
 52. The compoundof claim 31 wherein A or D is nitrogen.
 53. The compound of claim 31wherein m is
 2. 54. The compound of claim 31 wherein one of R* or R** issubstituted phenyl substituted with 1 to 5 substituents each of which isindependently selected from alkyl (greater than C₆), aryl, substitutedaryl, benzyl, substituted benzyl, (CH₂)_(k)OH (where k=1, 2 or 3),CH₂NH₂, CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H, CO₂alkyl, CONH₂, CONHalkyl,CON(alkyl)₂, OH, alkoxy (greater than C₆), aryloxy, SH, S-alkyl, S-aryl,SO₂alkyl, SO₃H, SCF₃, CN, NO₂, NH₂, 2^(o)-amino, 3^(o)-amino, NH₂SO₂,OCH₂CH₂NH₂, OCH₂CH₂NHalkyl, OCH₂CH₂N(alkyl)₂, oxazolidin-2-yl, and alkylsubstituted oxazolidin-2-yl.
 55. The compound of claim 44 wherein saidphenyl has the structure

wherein R⁴, R⁵, R⁶, R⁷ and R⁸ are each independently selected from thegroup consisting of hydrogen, alkyl, cycloalkyl, aryl, substituted aryl,aralkyl, substituted aralkyl, (CH₂)_(k)OH (where k=1, 2 or 3), CH₂NH₂,CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H, CO₂alkyl, CONH₂, CONHalkyl,CON(alkyl)₂, OH, alkoxy, aryloxy, SH, S-alkyl, S-aryl, SO₂alkyl,SO₂N(alkyl)₂, SO₂NHalkyl, SO₂NH₂, SO₃H, SCF₃, CN, halogen, CF₃, NO₂,NH₂, 2^(o)-amino, 3^(o)-amino, NH₂SO₂, OCH₂CH₂NH₂, OCH₂CH₂NHalkyl,OCH₂CH₂N(alkyl)₂, oxazolidin-2-yl, or alkyl substituted oxazolidin-2-yl56. The compound of claim 31 wherein R* and R** are each3-chloro-4-methylphenyl, R⁹ is H, R¹¹ is H, z is 1, A is CH, D is CH, Eis OH, m=0, r is 1, and G is ═O (double bonded oxygen).
 57. The compoundof claim 31 wherein R* and R** are each 2-methyl-4-chlorophenyl, R⁹ isH, R¹¹ is H, z is 1, A is CH, D is CH, E is OH, m=0, r is 1, and G is ═O(double bonded oxygen).
 58. The compound of claim 31 wherein one of R*and R** is a substituted or unsubstituted aralkyl.
 59. The compound ofclaim 31 wherein R* and R** are each a substituted or unsubstitutedaralkyl.
 60. The compound of claim 58 or 59 wherein said benzyl has thestructure

wherein R⁴, R⁵, R⁶, R⁷ and R⁸ are each independently selected from thegroup consisting of alkyl, cycloalkyl, aryl, substituted aryl, aralkyl,substituted aralkyl, (CH₂)_(k)OH (where k=1, 2 or 3), CH₂NH₂,CH₂NH-alkyl, CH₂N(alkyl)₂, CO₂H, CO₂alkyl, CONH₂, CONHalkyl,CON(alkyl)₂, OH, alkoxy, aryloxy, SH, S-alkyl, S-aryl, SO₂alkyl,SO₂N(alkyl)₂, SO₂NHalkyl, SO₂NH₂, SO₃H, SCF₃, CN, halogen, CF₃, NO₂,NH₂, 2^(o)-amino, 3^(o)-amino, NH₂SO₂, OCH₂CH₂NH₂, OCH₂CH₂NHalkyl,OCH₂CH₂N(alkyl)₂, oxazolidin-2-yl, or alkyl substituted oxazolidin-2-yl.61. The compound of claim 31 wherein one of R* and R** is a substitutedor unsubstituted heterocycle.
 62. The compound of claim 31 wherein R*and R** are each a substituted or unsubstituted heterocycle.
 63. Thecompound of claim 61 and 62 wherein said heterocycle has the structure

wherein X═CH═CH, N═CH, NR¹³ (wherein R¹³═H, alkyl, aryl or aralkyl), O,or S and wherein Y═CH or N and wherein R¹, R², and R³ are eachindependently selected from the group consisting of hydrogen, alkyl,cycloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl,(CH₂)_(k)OH (where k=1, 2 or 3), CH₂NH₂, CH₂NH-alkyl, CH₂N(alkyl)₂,CO₂H, CO₂alkyl, CONH₂, S-alkyl, S-aryl, SO₂alkyl, SO₂N(alkyl)₂,SO₂NHalkyl, SO₂NH₂, SO₃H, SCF₃, CN, halogen, CF₃, NO₂, oxazolidin-2-yl,or alkyl substituted oxazolidin-2-yl.
 64. A compound of claim 31 whereinR⁹ is H, R¹¹ is H, z is 1, A is CH, D is CH, E is OH, m=0, r is 1, and Gis ═O (double bonded oxygen)
 65. A compound of claim 31 furthercomprising a solvent adduct having the structure

and wherein R*** is H or alkyl.
 66. A compound having the structure of acompound of Table
 1. 67. A compound having the structure of a compoundof Table
 2. 68. A compound having the structure of a compound of Table3.
 69. A compound having the structure of a compound of Table
 4. 70. Acomposition comprising a compound of claim 1 in a pharmaceuticallyacceptable carrier.
 71. A method for treating a microbial-caused diseasein a patient afflicted therewith comprising administering to saidpatient a therapeutically effective amount of a compound of claim 1 to69.
 72. The method of claim 71 wherein said microbe is a bacterium. 73.The method of claim 72 wherein said bacterium is a gram positivebacterium.
 74. The method of claim 73 wherein said gram positivebacterium is a member selected from the group consisting ofStaphylococcus species, Streptococcus species, Bacillus species,Mycobacterium species, Corynebacterium species, Clostridium species,Actinomyces species, Enterococcus species, and Streptomyces species; 75.The method of claim 72 wherein said bacterium is a gram negativebacterium.
 76. The method of claim 75 wherein said gram negativebacterium is a member selected from the group consisting ofAcinetobacter species, Neisseria species, Pseudomonas species, Brucellaspecies, Agrobacterium species, Bordetella species, Escherichia species,Shigella species, Yersinia species, Salmonella species, Klebsiellaspecies, Enterobacter species, Haemophilus species, Pasteurella species,Streptobacillus species, spirochetal species, Campylobacter species,Vibrio species, and Helicobacter species.
 77. The method of claim 72wherein said bacterium is a member selected from the group consisting ofStaphylococcus aureus; Staphylococcus epidermidis, Staphylococcussaprophyticus; Streptococcus pyogenes; Streptococcus agalactiae;Streptococcus pneumoniae; Enterococcus faecalis; Enterococcus faecium;Bacillus anthracis; Mycobacterium avium; Mycobacterium tuberculosis,Acinetobacter baumanii; Corynebacterium diphtheria; Clostridiumperfringens; Clostridium botulinum; Clostridium tetani; Neisseriagonorrhoeae; Neisseria meningitidis; Pseudomonas aeruginosa; Legionellapneumophila; Escherichia coli; Yersinia pestis; Haemophilus influenzae;Helicobacter pylon; Campylobacter fetus; Campylobacter jejuni; Vibriocholerae; Vibrio parahemolyticus; Trepomena pallidum; Actinomycesisraelii; Rickettsia prowazekii; Rickettsia rickettsii; Chlamydiatrachomatis; Chlamydia psittaci; Brucella abortus; Agrobacteriumtumefaciens; and Francisella tularensis.
 78. The method of claim 71wherein said microbe is a fungus or yeast
 79. The method of claim 78wherein said yeast is a member selected from the group consisting ofCandida albicans, Candida glabrata, Candida krusei, Candida tropicalis,or Candida parapsilosis.
 80. The method of claim 78 wherein said fungusis a member selected from the group consisting of Aspergillus species,Trichophyton species, Microsporium species, Cryptococcus neoformans,Blastomyces dermatitidis, Coccidiodes immitis, Histoplasma capsulatum,or Paracoccidioides brasiliensis.